EZLabManual

-1-

EZLabManual INTRODUCTION This manual summarizes policies for employees and partners/owners of EZ Lab Diagnostic&LaboratoryClinic. This is a tool to help promote a cooperative and healthy atmosphere, to spell out policies relative to hours, wages, conditions of employment and to provide for the administrationofthesepoliciesintheinterestsofallconcerned,inkeepingwithconditionsin ourareaandindustry. We are presenting this employee handbook because we feel that if you understand basicallywhatisexpectedofyou,andwhatyoumayexpectofthecompany,weshallhavean organizationwhichbettermeetstheneedsofourcustomers. Thestatementsassetforthinthisbookhavenotbeenarbitrarilyestablished.Eachof them has a sound background of common sense based on the experiences of this company. Employeeshavesuggestedmanyandwewillfurtherwelcomesuggestionsfromyouthatwill aidinmaintainingaconstructiveandharmoniousrelationship. At the time new policies or revisions are issued, they shall be incorporated into the manual.Thecompanywillpostnotificationsofchangestothepoliciesastheyoccur. In order to ensure that policy interpretations are consistent, the Office of the Human Resources or designee is solely responsible for interpreting policy application and intent. His/herinterpretationsshallbeapplicabletoallpertinentsituations. TheManualshallapplytoallemployeesandpartnersfromthetimeofitsadoption.The companymayalter,eliminate,oraddtoanyoftheprovisionsoftheManualatanytimeandfor any reason, and such alterations, eliminations, or additions shall apply to all employees and partnersfromthetimeoftheirpublication. Our single most common goal must be to work together to meet the needs of our clients,rememberingourclientsaremutuallyourmostimportantasset.

-2-

THE COMPANY: Orientation, Mission and Vision

EZLabManual

FOREWORD

EZ Lab Diagnostic and Laboratory Clinic is a nonstock, nonprofit diagnostic and laboratory clinical laboratory offering leading edge testing services in the areas of clinical trials and toxicology,allunderoneroofatitsstateoftheartfacility. Strategicallylocatedat#46QuezonAvenue,Brgy.1,LucenaCity,EZlabisacomprehensiveand privately held laboratory operating on a provincial basis, delivering a quick, quality and convenientserviceatcompetitiveprices. EZ Laboratory also offers mobile services for more convenience of our clients. Those with abnormalfindingscanavailofmedicalconsultation,atdiscountedrates,withourdoctorswho are specialists and subspecialist in Cardiology, Nephrology, Pulmonology, Gastroentrology, Surgery,ObstetricGynecology,andOtolaryngology. To assure of the accuracy of our results, all examinations are performed with utmost ability, using high quality materials at the shortest possible time. That is the reason why we have devotedareputationasapremierlaboratory,especiallyintheareaofQuezon.Weevenoffera moneybackguaranteetoanyprovenerroneoustestresult. MISSION EZLabDiagnosticandLaboratoryClinicsmissionistoprovideacompetitiveadvantageandfast service for its clients. They are dedicated to personalized service, the most rapid turnaround time in the industry in Quezon, innovative information management systems, and accurate testingtechnology. VISION EZ Lab Diagnostic and Laboratory Clinics vision is to be the undisputed leader in providing customized health information services to selected markets in Quezon province in particular andthePhilippinesingeneral. GOAL EZLabDiagnosticandLaboratoryClinicsmaingoalistobetheundisputedleaderinproviding customizedhealthinformationservicestoselectedmarketsintheprovince.

To conduct scientific, evidencebased research from bench research to patientcare, evaluating pharmacokinetics, pharmacodynamics, bioavailability, efficacy, clinical application,andsafety. Todevelopinnovativebiosciencesresearchmethodologiestoevaluatefromrandomized controlledtrialstowholepracticeoutcomesbasedresearch.

-3-

EZLabManual

Toestablishsystemsmetricsofbiomedicaloutcomesmarkersandnaturaltherapiesfor preventionandhealthwellnessprograms. To disseminate clinically valuable information and scientific research about issues in NaturopathicMedicinethatimpactsthehealthcaresystem.

STRATEGY

CompensationandbenefitpackageswillsupporttheLaboratorysefforttorecruitand retain a highperforming and diverse work force. The laboratory will provide a high level of service to its customers by making use of available knowledge, skills and technology and throughfacetofaceinteractionwiththosecustomers.AllEZlabstaffandpartnerswillfillan advocacyrole,whenappropriate.Infillingthisadvocacyrole,theywillbemindfuloftheneed tobalanceemployeerightsagainstthelegitimatebusinessneedsofthecompany. Managementsponsoredtraininganddevelopmentactivitieswillbeprovidedto enablesupervisorsandemployeestoimprovetheirskillsandabilitiesincarrying outtheirpresentrolesandensuresafeandeffectiveworkandhealthhabits. HR staff will provide appropriate career development opportunities to enable employeestobetterthemselves. The HR and Management will carry out Laboratory personnel policies and proceduresinanopenandhonestfashion,welcominginputandadvicefromour customers. The laboratory personnel will partner with upper management by providing professional and expert and advice and services on matters that impact upon personnelissues.

-4-

COMPANYPOLICIES:Values
VALUES

EZLabManual

BECAUSEOFOUR CLIENTFOCUS COMMITMENT COMPASSION FLEXIBILITY INTEGRITY PERSEVERANCE QUALITY WEAREENDEAVORTO Fulfillourclients'individualneeds,creatingcustomized solutionsandprovidingthemacompetitiveadvantage. Dedicateourselvestohelpingeachotherdoourbest Begenerous,empathic,Considerateandkindinourinteractions withothers Beopenandaccommodatinginouractionsanddecisions Adheretoacodeofethicsthatpromotesconfidenceandtrust Staythecourseuntilweachievesuccess. Exhibitandexceedthehigheststandardsofexcellenceinevery aspectofourlives Exhibitandexceedthehigheststandardsofexcellenceinevery aspectofourlives

QUALITY SERVICETOHUMANITY Enhance lives through scientific contributions, technology, and gooddeeds,withoutconsideringselfinterests. TEAMWORK Work together to achieve common goals and a meaningful purposeinourlives,andthelivesofothers. -5-

EZLabManual

THEWORKFORCE:JOB DESCRIPTIONAND SPECIFICATION

-6-

JOBDESCRIPTIONANDSPECIFICATION GENERALMANAGER

EZLabManual

DUTIESANDRESPONSIBILITIES: 1. TheprimarydutyoftheGeneralManageristobethedirectrepresentativecapableof makingsounddecisionsonbehalfofPartners. 2. Administer the affairs of the partnership in accordance with organizational policies. Ensure the maintenance of official records, bylaws, and standing rules according to ManagementCommitteeaction. 3. Work closely with the Partners and leadership of the organized Medical Staff in developingthestrategicdirectionandmajorpoliciesoftheinstitution.Consultwiththe Partnersonmattersconcerningmemberissuesandconcerns. 4. Attend Partnership and Coordinating Committee meetings, disseminating information betweengovernancebodiesandstaff,andreportingonworkplaceoperations,finances, planning, and other matters as necessary. Ensure representation at conferences and keep the Management Committee, staff, and members informed of EZ Lab and other relevant issues and activities. Promote effective communication on all levels of the organization. 5. Researchandwritediscussionpapers,analysisdocumentsandproposalsasneededto assist the organization in determining and oversee preparation of an Annual Report summarizingprogressonshortandlongrangeplansandgoals.Provideleadershipand vision to the organization by assisting the Management Committee and staff with the developmentoflongrangeandannualplans,andwiththeevaluationandreportingof progressonplans. 6. Be available for employees that experience work and/or personal problems providing appropriate coaching, counseling, direction and resolution. Proactively provide information, history, continuity, support and guidance to all governance bodies and members,inconjunctionwiththestaffteam. 7. Respondtoandresolveemployeerelationsissuesexpressedbyteammembers;create and maintain high quality work environment so team members are motivated to perform at their highest level. Use appropriate judgment in upward communication regardingdepartmentoremployeeconcerns. 8. Address disciplinary and/or performance problems according to company policy. Preparewarningsandcommunicateeffectivelywithemployeesonwarningsandmake effective/appropriatedecisionsrelativetocorrectiveaction. 9. The General Manager will recruit, hire and train other personnel for permitted areas and resolve personnel problems, will handle complaints, infractions, etc., which are referredtohim/herandtakenecessaryactiontoresolvethesesituations. 10. Ensure that all staff members receive appropriate training to perform their jobs effectively. 11. Overseeadministrationofstaffbenefits.

-7-

EZLabManual 12. Be a resource for employee and guest conflict resolution. Responsible for suggesting methods to improve area operations, efficiency and service to both internal and externalcustomers. 13. Actasarbitrator/mediatorinworkandpersonnel/personaldisputes. 14. Overseethemaintenanceofpersonnelandotheradministrativerecords. 15. Provide vision and oversight for the technology needs of the organization. Oversee ongoing, short range and long range property management to ensure the viability, continuityandgrowthofEZLabassets. 16. Assuringthesoundfiscaloperationofthelaboratorywhilepromotingservicesthatare produced in a costeffective manner. Analyzes and controls expenditures of the departmenttoconformtobudgetaryrequirements. 17. Responsible for managing the daytoday operations of the laboratoryand its entities. Establishingasystemforassuringthathighqualityisprovided. JOBSPECIFICATION
1. Masters Degree in laboratory administration, public health business administration or management,orrelatedfieldisrequired.

2. With experience in business management, planning, financial oversight, personnel management,includinghiring,supervision,evaluationandbenefitsadministration.With experienceworkingwithapartnersandcommittees. 3. Ability to read, analyzes, and interprets general business periodicals, professional journals,technicalprocedures,orgovernmentalregulations;towritereports,business correspondence,andproceduremanuals;tointerpretavarietyofinstructionsfurnished inwritten,oral,diagram,orscheduleform. 4. Ability to communicate effectively, both orally and in writing. Ability to effectively present information and respond to questions from groups of managers, clients, customers, and the general public; to interact, relate to, work with, and support the activities of highly qualified, dynamic physicians as individuals and members of the organizedMedicalStaff. 5. Demonstrated ability to manage and supervise a staff team. Ability to establish and maintain effective working relationships with staff and the general public using tact, courtesy,goodjudgment,mediaandpublicrelations. 6. Proficiencywithofficecomputerequipmentandsoftware 7. Ability to solve practical problems and deal with a variety of concrete variables in situationswhereonlylimitedstandardizationexists. 8. Abilitytoorganizeandprioritizeavarietyoftasksandprojects. 9. Demonstratedabilitytoshareskillsandknowledgewithothers.

-8-

JOBDESCRIPTIONANDSPECIFICATION OFFICEMANAGER SUMMARY:

EZLabManual

ReportsdirectlytotheGeneralManagerandmaintainsacloseworkingrelationshipwith administrative and other personnel. The office manager has both clinical and management responsibilities. S/He serves as manager, supervising personnel and developing, coordinating andmanagingofthebusiness. DUTIESANDRESPONSIBILITIES: 1. Prepareandmaintainallgoverningbodyminutes,practicerecordsandotherdata. 2. Monitoremployeestimeslips(hoursworked,vacationleave,sickleave,etc.). 3. Maintain payroll to include maintenance of employee record, computation of federal and state payroll taxes, SSS, Philhealth, PagIbig Contribution and miscellaneous deductions,etc.,asdirected. 4. Matchinginvoicestopackingslipsandpurchaseorders. 5. Processandverifypaymentofinvoicesonatimelybasis 6. Verificationofinvoicesreceivedforquantity,unitprice,extensions,anddiscounts. 7. Preparemonthlytotalsofinvoices,accountspayable,cashdisbursements,etc.,asmay bedirected. 8. Forwardinvoicestoauditor,generalmanagersandfinancecommitteeforapprovalfor payment. 9. Sendchecksandmaintainsafileofpaidinvoices. 10. TheOfficeManagerwillalsoprovideclericalsupporttootherpersonnel. 11. Evaluates,counselsandpromotesthegrowthanddevelopmentoftheclinicalstaff 12. Dealfairlyandconsistentlywitheachemployee. 13. Beaccessibleto,andasoundingboardfor,allemployees. 14. Developandrecommendwrittenpersonnelpoliciesforvacation,sickleave,employee benefitsandrelatedmatters. 15. Conductperiodicreviewsandevaluationsofallemployees. 16. Developandimplementemployeetrainingprograms. 17. Establishandreviseasneededjobdescriptionsandproceduresmanuals. 18. PlanandScheduleofficestaffworkassignments. 19. KeeprecordsonallpersonnelofEZLabDiagnostic&MultiSpecialistCenter 20. S/HeservesassitemanagerandisdesignatedasactingadministratorwhentheOfficer oftheMonthisnotonsite.

-9-

EZLabManual 21. Work closely with the company partners, owner, general managers to meet the companysneed. 22. Consults with and advises the General Managers regarding needs, goals, future plans andproblemspertinenttoclinicalprograms. 23. CollaborateswiththeGeneralManagerstoestablishandmaintaindirection,operation practices, procedures, and standards for the organization, operation and performance pertinenttothepatient. 24. DirecttheoverallBusinessOfficeactivities. 25. Coordinate and communicate information and activities among staff, committees, ownersandanyothergroupswithintheorganizationalstructureofEZLab. 26. Assures that professional and support staff properly implement clinical policies and procedures 27. Receivesvisitorstoofficeandrespondstoneeds. 28. Recommendtothegeneralmanagermaintenanceneedsforallofficeequipment. 29. Do everything possible to become a proficient computer operator. Supervise all computerrelated procedures. Plan overall utilization of computer capabilities. Responsible to see that all changes are kept uptodate. Generate reports when requiredbygeneralmanagersorotherownerofthebusiness. 30. Provides an uninterrupted support of total office operations. Orders nonmedical suppliesasneeded. 31. Maintainpersonnelfilesonallpersonnel,ownersandsupport/officestaff,asoutlinedin EmployeePolicy. 32. Maintainrecordsandfollowuponitemsborrowedfromtheoffice,andthecollectionof moneyowedtothelaboratory(i.e.,personallongdistancecalls,wholesalepurchasing cards,etc.). 33. MaintainanOfficeProcedureManualsothatinformationastothefunctionoftheoffice iskeptuptodateandiseasilyaccessibletoothersshoulditbeneeded. 34. Asnecessary,updateanyprintedmaterialssothatinformationisaccurateandcurrent. 35. Performallotherduties,asassigned

- 10 -

EZLabManual JOBSPECIFICATION: 1. Degree in business or accounting and two years of related office and health care environment experience, (one year of supervisory experience) or an equivalent education/experience. 2. Mustbecomputerliterate.ProficientuseinMicrosoftWord,ExcelandAccess. 3. Mustpossesstheabilitytoexamineandverifyfinancialdocumentsandreports. 4. Must be able to prepare financial and other records in a systematic, neat, and legible manner. 5. Must possess familiarity with clinical operating policies and procedures, insurance industrypractices,humanresourcemanagementandfinancial/accountingprinciples. 6. Abilitytoprofessionallyrepresentthephysicians. 7. Excellentverbalandwrittencommunicationskills. 8. Experiencewithcustomerservice. 9. Strongorganizationalandinterpersonalskills. 10. Abilitytofulfillleadershiprole. 11. Abilitytocreatechangeasneeded. 12. Abilitytoprioritizeresponsibilities. 13. Mustbeabletoworkindependentlyandwithoutsupervision. 14. Abilitytomaintainconfidentialityofsensitiveinformation. 15. Abilitytomultitaskefficientlyandeffectively. 16. Takesinitiativeasnecessary. 17. Abilitytoactcalmlyandeffectivelyinabusyorstressfulsituation. 18. Requiresadherencetoallpoliciesandprocedures. 19. Mustbeabletoestablishandmaintaineffectiveworkingrelationshipswithphysicians, managersandpeers. 20. Must be able to reason, analyze, prioritize, conceptualize, make judgments, and solve problems.

- 11 -

EZLabManual RECEPTIONIST SUMMARY: This position is responsible for the professional and efficient managing of patients, visitors,clients,telephonecallsandmessages,thenassistinclericalneeds,aswellasavariety ofadministrativeandotherdutiesthatsupportconsumerservices,informationandreferraland theoperationandpresentationofaprofessionaloffice. DUTIESANDRESPONSIBILITIES: FrontDeskReception 1. Greetpatientsuponarrival. 2. Receivepatientsattheclinicaswalkin/followup/referral. 3. Readanddeterminepatients'needsbyreadingrequestsorinstructionsfromphysician andexplaintopatient. 4. Fill up the request form of EZ Lab by asking patient about his/her complete name, birthday,ageandsex.Checktheappropriateboxrequestedbythephysicianandbythe patient. 5. Determineseligibilityforpatientsabilitytopay.OffertheEZLabdiscountlikesenior citizen,loyaltycardandotherdiscounts. 6. Receivepaymentsandissuingofpatientsreceipts. 7. SubmittherequestformtoLaboratoryAssistant. 8. Typingandprintingoflaboratoryandxrayresult. 9. Releasingoflaboratoryresults. 10. Postallcashonadailybasis. 11. Toberesponsibledailycashandcheckrespectofconsultationfeesanddailycollections. 12. Reportandrecordcashcollectionsandexpensesincashbook.Verifyaccuracyofdaily depositslipwithcashreceiptsjournal. 13. Maintainsacompleteandsystematicsetofrecordsofbusinesstransactions.Balances books and prepares reports to show receipts, expenditures, accounts payable, and variousotheritems. 14. Recordandcomputereferralscommission. 15. Maintainthemedicalrecordsfunctionsthroughpulling,filing,assemblingandretrieving patientrecords. 16. Workingcloselywiththeclinicassistant,ensurethesmoothoperationofclinicalareas, includingpatientflow,processingofmedicalrecords,andphysicianscheduling. 17. Oversee the practice computer system; work closely with consultants to maintain current,effectivehardwareandsoftwareensuredatabackupprocedures,andmaintain thepracticewebsite. - 12 -

EZLabManual 18. Answersincomingcallsanddirectscallscorrectly. 19. Takes&deliversmessages 20. Checksfaxmachineforincomingfaxes&deliverstoappropriatepersonnel. 21. Providesappropriateinformationandreferralwhennecessary/requested. 22. Ensuresthatcommonareasareequippedwithofficesuppliesasneeded. 23. Assist in forwarding patient files, letters and/or results to other medical centre as requested,ensuringappropriatedocumentationoftransfersarekept. 24. Scheduleappointmentsforpatientsaccordingtoestablishedprocedures 25. Sortsandfilescorrespondenceanddocumentsalphabetically,numerically,orbyother establishedclassifications;removesandreturnsmaterialfromfiles;preparesfilefolders bysubject,nameorotherheadings. 26. Performsotherrelatedworkasassigned. JOBSPECIFICATION: 1. GraduateofBusinessAdministration 2. Onetotwoyearsmedicalofficeexperiencepreferred. 3. Successful candidate must have excellent attention to detail, the ability to follow instructionspreciselyandunderstandallconfidentialitypoliciesandproceduresrelative tomedicalrecords. 4. Havewordprocessingexperienceandgoodorganizationalskills.Specialskillsdesiredfor thispositionincludeknowledgeofMicrosoftWord,ExcelandAccess. 5. Excellent customer service and phone/communication skills, strong organizational abilitiesandthecapacitytoworkindependently. 6. Knowledgeofcomplianceandcostreimbursement. 7. Maintainsconfidentialityofpatientinformation. 8. Abilitytoprocesspatientsandagencyinquiries. 9. Pleasantphonevoicewithpropergrammarspoken 10. Abilitytoworkeffectivelyinafastpacedofficeenvironmentisessential. 11. Workenvironmentmaturityandprofessionalism. 12. Abilitytowritesimplecorrespondence. 13. Ability to effectively present information in oneonone and small group situations to customers,clients,andotheremployeesoftheorganization. 14. Ability to add, subtracts, multiply, and divides in all units of measure, using whole numbers,commonfractions,anddecimals.Abilitytocomputerate,ratio,andpercent andtodrawandinterpretbargraphs. 15. Experienceincashhandlingandsimpleaccountmanagement. 16. Abilitytoworkwellunderpressure 17. Abilitytoworkaccuratelyandefficiently 18. Knowledgeofpatientbillingprocedures 19. Someknowledgeofmodernofficepractices,proceduresandequipment. 20. SomeknowledgeofbusinessEnglish,spellingandarithmetic. - 13 -

JOBDESCRIPTIONANDSPECIFICATION

EZLabManual

21. Skill in establishing and maintaining effective working relationships with other employees,patients,clients,andthepublic.

MARKETINGMANAGER SUMMARY

Developsandimplementsstrategicmarketingplanforanorganization.Staysabreastof changes in the marketing environment to best serve the objectives of the organization and adjusts plans accordingly. Researches and develops pricing policies and recommends appropriatesaleschannels.Reliesonexperienceandjudgmenttoplanandaccomplishgoals. Typicallyreportstomanagementcommittee. DUTIESANDRESPONSIBILITIES: 1. Makealistoftargetmarket. 2. Prepare marketing strategic plan which will appropriate to target market and will benefitstheEZLab. 3. Researches, evaluates prepares and/or presents customized presentations for prospectivecompanieswhichhighlightbusinessadvantagestootherlaboratories. 4. Make a presentation or proposal and present to management committee for the approval. 5. Dealswithclientsaboutourproductsandservices. 6. Providescompaniesandcommunitieswithinformationrelatingtocompetitors. 7. Researches, evaluates and provides interested businesses with information on trade opportunities,potentialmarketsandcustomers. 8. Schedules and facilitates meetings and site visits for companies, clients, government officialsanddignitariesexploringbusinessopportunities. 9. Participates in the business negotiation process; assists with the completion of necessarypaperwork. 10. Plansandschedulesworkactivitiestoachieveestablishedgoals. 11. Follow up target market and responsible to have a memorandum of agreement betweenEZlabandclient. 12. Preparesandsubmitsactivityreports. 13. Informs EZ Lab personnel about the schedule of his/her clients for laboratory examination. 14. Exercises independence and initiative in the performance of assigned responsibilities; receivesgeneraladministrativedirection. 15. Performsotherrelatedworkasassigned.

- 14 -

EZLabManual JOBSPECIFICATION 1. 2. 3. 4. BachelorinBusinessAdministrationMajorinMarketing. Twoormoreyearsofprofessionalexperienceinmarketingoflaboratoryservices. Withpleasingpersonality. IntroductoryknowledgeofbusinessfactorsaffectingthemarketingofEZLabproducts andservices,includingtradeleadsources,directandindirectmarketing,transportation, pricequotation,dutyrequirementsandproductclearance. Ability to prepare and present marketing and related promotional material to businesses. Abilitytopreparecompleteandconcisereports. Abilitytoestablishandmaintaineffectiveworkingrelationshipswithotheremployees, businessandtradeassociationrepresentatives,federal,stateandlocalofficials,foreign customersandgovernments,communityleadersandthegeneralpublic. Abilitytotravelextensivelyintheperformanceofassignedduties. Abilitytocommunicateproficientlyinadesignatedforeignlanguageasmayberequired fortheassignedprogramarea.

5. 6. 7.

8. 9.

XRAYTECHNOLOGIST

DUTIESANDRESPONSIBILITIES: 1. Determinepatients'xrayneedsbyreadingrequestsorinstructionsfromphysicianand explaintopatient. 2. Determinewhichscreenfilmcombinationwillneed. 3. Determinewhatsizeofxrayfilmandradiographiccassettewillneed. 4. Preparetheradiographicroomorexposureroom. 5. Dresspatientcorrectly. 6. Explaintothepatientwhatwillbedoingandwhatisexpectedofthem. 7. Arrangespatientsforradiologicalexaminationswithcomfort. 8. Positionpatientonexaminingtable,setupandadjustequipmenttoviewspecificbody areaasrequestedbyphysician. 9. Performproceduresusingxrayequipmentundersupervisionofphysician. 10. Preventspatientfrombeingexposedtounnecessaryradiation. 11. Measuretheparttoberadiographed. 12. Determinetheradiographictechniquetobeusedandsetonthexraymachine. 13. Use beamrestrictive devices and patientshielding techniques to minimize radiation exposuretopatientandstaff. 14. Collimatedthebeamsothattheonlyareaofinterestisincluded. 15. Identifyrightandleftofthepatientwithproperleadmarker.

- 15 -

EZLabManual 16. Explainthatyouaregoingtodevelopandviewthefilmshavebeentakentodetermine ifadequateradiographshavebeen. 17. Record the date, time number of films, name of patient and patient history on the requisition. 18. Properlycritiqueradiograph.Ifnoradiographneedstoberepeated,placeinproperslot orsendwithproperifindicated. 19. Assistthepatient. 20. Xray techs must observe proper safety procedures to avoid unnecessary exposure to harmfulradiation. 21. Processexposedradiographsusingfilmprocessorsorcomputergeneratedmethods. 22. Producesradiographs(xrays)toaidinthediagnosisofmedicalproblems. 23. Operate the testing equipment, develop xray films, and pass them along to the Radiologistfordiagnosis. JOBSPECIFICATION: 1. Healthcare Professionals who primarily take Xrays and Administer Nonradioactive materials 2. CollegePreferablyRadiographicTechnologist/XrayTechnologist 3. AtleasttwoyearsofexperienceasaRadiologicTechnologist. 4. CurrentPRClicensewithDiplomaandTranscripts. 5. Has knowledge of commonly used concepts, practices, and procedures within a particularfield. 6. Reliesoninstructionsandpreestablishedguidelinestoperformthefunctionsofthejob. 7. Acertaindegreeofcreativityandlatitudeisrequired. 8. Worksunderimmediatesupervision. CLINICASSISTANT

DUTIESANDRESPONSIBILITIES: 1. Assistsincheckingvitalsigns,heightandweightasassigned. 2. Arrange bedside unit, clinic room and equipment; assists physician with washing and sterilizingequipment. 3. Performsrelatedworkasassigned. JOBSPECIFICATION: 1. Oneormoreyearsofexperienceasalaboratory&clinicassistant.

- 16 -

EZLabManual 2. Introductoryknowledgeofdirectcareproceduresandtechniquesinvolvedinproviding basiccare. 3. Abilitytoprepareandmaintainrecords,chartsandreports. 4. Abilitytomaintainpatienceandtactindealingwithpatients. 5. Abilitytotakevitalsigns. 6. Abilitytofollowdirections. 7. Abilitytocompleteabasiccourseofpatientdirectcareinstruction. 8. Ability to establish and maintain effective working relationships with staff and management. 9. Introductoryknowledgeofmedicalterminology. LABORATORYAIDE JOBDESCRIPTION 1. The main duty of the laboratory assistant will be to help with the setup for weekly laboratories.Cleaninguppreviouslabs,andputtingtogetheranyequipmentthatwillbe needed. 2. Assistswithreceiptandinspectionofrawmaterials. 3. Miscellaneousdutiesmayberequestedastheneedarises. 4. Workisperformedunderthedirectsupervisionofahigherlevellaboratoryemployee. 5. Receivingoflaboratorysuppliesandrecordingintheledger. 6. Maintainsavarietyoflaboratoryrecordsandfiles. 7. Keeprecordsofchemicals,apparatusandotherglasswares. 8. Returnsunusedchemicalstodesignatedcabinetsandcleansworkarea. 9. Check incoming specimens for proper identification and sufficient quantity, record patientsname,age,sex,dateofbirth.Attachedlabrequesttosamples. 10. Recorddailytotalsofproceduresperformed. JOBSPECIFICATION: 1. College,withmedicalbackground. 2. AbilitytooperateECGMachine. 3. Phlebotomist 4. Willingnesstoperformrespectivelaboratorytasks; 5. Abilitytokeepaccuraterecordsandmakesimplereports; 6. Theworkstudyshouldhavegoodorganizationalandcommunicationskills 7. Abilitytodealeffectivelywithstaffandpatients 8. Physicalconditioncommensuratewiththedemandsoftheposition. - 17

EZLabManual MEDICALTECHNICIAN SUMMARY: Performvariouslaboratoryproceduresasenumeratedunderdutiesandresponsibilities ofMedicalTechnologist. DUTIESANDRESPONSIBILITIES: 1. Drawsbloodandcollectsurine,stool,sputumandothersamplesforanalysisasordered byphysicians. 2. Examines stained blood smears microscopically and refers any abnormal cells to superiors. 3. Performs chemical analysis. Analyses human material or other products submitted to the laboratory, using photometric, calorimetric, titrimetric, radioisotope, or any other chemicalorphysicalproceduresapplicabletoclinicalchemistry. 4. Maintainsbasicrecordsconsistentwithassignedresponsibilities. 5. Maintainsaccuraterecordsandinitiatescorrectiveactionwhennecessary. 6. Cleansandmaintainssamplecontainers,laboratoryequipmentandworkareas. 7. Assistsinallareasofthedailyoperationofthelaboratory. 8. Guaranteesconfidentialityoflaboratorytestresultsandpatientidentity. 9. Performsdailymaintenanceonanalyzersandotherlaboratoryequipment. 10. May perform and report microbiological, serological and chemical analysis on clinical samples. 11. Mayperformandreporthematologicalandchemicalexaminationsonbloodandother bodyfluidsforthedetectionandidentificationofcommunicableandotherdiseases. 12. Assistwithcollectionandanalysisoflabdata. 13. Requestandrecommendstheorderingofneededreagents,glasswaresandequipments totheGeneralManager 14. Purchase reagents and other need in the laboratory upon approval of the General Manager 15. Monitors and ensure that all necessary supplies and equipments in the laboratory are availableandoperational. 16. Responsible for keeping the laboratory equipments and apparatus in good working condition. 17. Seetoitthatallexaminationhavewrittenrequestbeforetheyaredone. 18. Responsibleforfollowingupofsendoutexaminationanditsresults. 19. Updatesrecordsoflaboratoryresultsinthelogbook. 20. InformtheGeneralManageraboutfaultyapparatusorequipmentsforpossiblerepair orreplacement. 21. Checkandrechecksalllaboratoryresults. 22. Performsvariouslaboratoryproceduressuchascollection,handlingandpreservationof specimen. - 18

EZLabManual 23. Performs blood chemistry procedures such as FBS, BUN, Uric Acid, Creatinine, Lipid Profile,SGOT,SGPT,CardiacEnzymes,etc. 24. PerformshematologicalproceduressuchasCBC,PlateletCount,ClottingTime,Bleeding Time,ESR,etc. 25. Performs serological and blood banking procedures such as blood typing, Rheumatoid Antigen,VDRL,RPR,ASO,etc. 26. Performs clinical microscopy and parasitological procedures such as routing urinalysis, fecalysis,pregnancytest,occultblood,seminalfluidanalysis,etc. 27. Reviewsprofessionalliterature 28. Operationalcharacteristicsofapparatus,equipmentandmaterialsusedinalaboratory. 29. Generalrulesforsafeexposureandhandlingofchemicalandbiologicalhazards. 30. Laboratorytechniquesandprocedures. 31. Practicesandprinciplesofbasiclaboratoryoperations. 32. Performsotherrelatedworkasassigned. JOBSPECIFICATION: 1. BachelorofScienceinMedicalTechnology. 2. Fiveormoreyearsasalaboratoryassistantperformingroutinechemical. 3. Basicknowledgeofchemical,biological,hematological,immunologicandmicrobiology laboratory and sampling principles, techniques, procedures, terminology and equipment. 4. Someknowledgeofbasicscienceterminology,conceptsandprinciples. 5. Someknowledgeoflaboratoryprocedures,techniquesandequipment. 6. Someknowledgeofblooddrawingtechniques. 7. Ability to properly operate microscopes, centrifuges, autoclaves, sterilizers or other laboratoryequipment. 8. Ability toapply propermethods of handling and disposing of chemicals and infectious materials. 9. Abilitytoperformassignedtasksaccordingtospecificinstructionsandclearlyprescribed procedures. 10. Abilitytokeeproutinelaboratoryrecords. 11. Ability to read, compared, identify and record laboratory data accurately, such as names,numbers,sampledescriptions,etc. 12. Abilitytoperformbasicmathematicsandmakeaccuratemeasurements. 13. Abilitytomakeaccurateobservationsandprepareaccuraterecordsoflaboratorytests. 14. Abilitytooperateautomatedanalyzersandperformsmanualtestsinaccordancewith detailedinstructions. 15. Ability to work with other employees, laboratory staff, health professionals and the generalpublic. - 19 -

EZLabManual 16. Workingknowledgeoflaboratorytestingtechniquesandprocedures. 17. Someknowledgeofbasicchemistry. 18. Skillintheuseandcareoflaboratoryequipment. 19. Abilitytoadheretooralandwritteninstructionsandtomakeaccurateobservations. 20. Ability to prepare accurate records and calculate results of laboratory tests, using standardequationsorformulas. 21. Abilitytoperceivecolorsnormally. 22. Abilitytodoworkthatrequiresmeticulousandthoroughperformance. MEDICALTECHNOLOGIST SUMMARY: Performmostanalyticaltestsonhumanbodyfluidsandtissuesprovidingaccuratedata tothehealthcareteamforoptimaldiagnosis,treatmentandmonitoringofdiseasestates. DUTIESANDRESPONSIBILITIES: 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. Performs coagulation studies on human blood and plasma. Performs chemical, macroscopic,andmicroscopicurinespecimenexaminations. Maintainsbasicrecordsconsistentwithassignedresponsibilities. Maintainsaccuraterecordsandinitiatescorrectiveactionwhennecessary. Cleansandmaintainssamplecontainers,laboratoryequipmentandworkareas. Assistsinallareasofthedailyoperationofthelaboratory. Guaranteesconfidentialityoflaboratorytestresultsandpatientidentity. Performsdailymaintenanceonanalyzersandotherlaboratoryequipment. May perform and report microbiological, serological and chemical analysis on clinical samples. Mayperformandreporthematologicalandchemicalexaminationsonbloodandother bodyfluidsforthedetectionandidentificationofcommunicableandotherdiseases. Assistwithcollectionandanalysisoflabdata. Requestandrecommendstheorderingofneededreagents,glasswaresandequipments totheGeneralManager Purchase reagents and other need in the laboratory upon approval of the General Manager Monitors and ensure that all necessary supplies and equipments in the laboratory are availableandoperational. Responsible for keeping the laboratory equipments and apparatus in good working condition. Seetoitthatallexaminationhavewrittenrequestbeforetheyaredone. - 20 -

EZLabManual 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. Responsibleforfollowingupofsendoutexaminationanditsresults. Updatesrecordsoflaboratoryresultsinthelogbook. InformtheGeneralManageraboutfaultyapparatusorequipmentsforpossiblerepair orreplacement. Checkandrechecksalllaboratoryresults. Performsvariouslaboratoryproceduressuchascollection,handlingandpreservationof specimen. Performs blood chemistry procedures such as FBS, BUN, Uric Acid, Creatinine, Lipid Profile,SGOT,SGPT,CardiacEnzymes,etc. PerformshematologicalproceduressuchasCBC,PlateletCount,ClottingTime,Bleeding Time,ESR,etc. Performs serological and blood banking procedures such as blood typing, Rheumatoid Antigen,VDRL,RPR,ASO,etc. Performs clinical microscopy and parasitological procedures such as routing urinalysis, fecalysis,pregnancytest,occultblood,seminalfluidanalysis,etc. Reviewsprofessionalliterature Operationalcharacteristicsofapparatus,equipmentandmaterialsusedinalaboratory. Generalrulesforsafeexposureandhandlingofchemicalandbiologicalhazards. Laboratorytechniquesandprocedures. Practicesandprinciplesofbasiclaboratoryoperations. Performsotherrelatedworkasassigned.

JOBSPECIFICATION 1. 2. 3. 4. RegisteredMedicaltechnologist LaboratoryDrugTestingAnalyst Fiveormoreyearsasalaboratoryassistantperformingroutinechemical. Basic knowledge of chemical, biological, hematological, immunologic and microbiology laboratory and sampling principles, techniques, procedures, terminologyandequipment. 5. Someknowledgeofbasicscienceterminology,conceptsandprinciples. 6. Someknowledgeoflaboratoryprocedures,techniquesandequipment. 7. Someknowledgeofblooddrawingtechniques. 8. Ability to properly operate microscopes, centrifuges, autoclaves, sterilizers or otherlaboratoryequipment. 9. Ability to apply proper methods of handling and disposing of chemicals and infectiousmaterials. 10. Ability to perform assigned tasks according to specific instructions and clearly prescribedprocedures. 11. Abilitytokeeproutinelaboratoryrecords.

- 21 -

EZLabManual 12. Abilitytoread,compared,identifyandrecordlaboratorydataaccurately,suchas names,numbers,sampledescriptions,etc. 13. Abilitytoperformbasicmathematicsandmakeaccuratemeasurements. 14. Ability to make accurate observations and prepare accurate records of laboratorytests. 15. Abilitytooperateautomatedanalyzersandperformsmanualtestsinaccordance withdetailedinstructions. 16. Abilitytoworkwithotheremployees,laboratorystaff,healthprofessionalsand thegeneralpublic. 17. Workingknowledgeoflaboratorytestingtechniquesandprocedures. 18. Someknowledgeofbasicchemistry. 19. Skillintheuseandcareoflaboratoryequipment. 20. Ability to adhere to oral and written instructions and to make accurate observations. 21. Ability to prepare accurate records and calculate results of laboratory tests, usingstandardequationsorformulas. 22. Abilitytoperceivecolorsnormally. 23. Abilitytodoworkthatrequiresmeticulousandthoroughperformance. JANITOR

DUTIESANDRESPONSIBILITIES: 1. Open the building at the designated time and have all front desk activities fully operationalatthestartofbusinesshours. 2. Assure the readiness of the reception area for each working day including the receptionist table, front glass, water dispenser, trash can, disposable cup and other equipments. 3. Cleaningofclinicroomincludingchangingofblanket,pillowcaseandgown. 4. Cleaningoflaboratoryanddrugtestroomincludingtables,refrigeratorandcabinets. 5. Properlyarrangedthejanitorialequipments. 6. Sweepingandmoppingofthefloor. 7. Maintainsthecleanlinessofcomfortrooms. 8. Orderingofjanitorialsupplies. 9. Responsibleforthemaintenanceofairconditionandotherequipments. 10. Performsavarietyofminorbuildingmaintenanceandcleaningtasks. 11. Securethebuildingatthecloseofeachworkingday;turnofforunplugappliancesand machinesaccordingtocompanyprotocolsandlockallentrances - 22 -

EZLabManual 12. Worksunderimmediatesupervision.Typicallyreportstoamanager. 13. Ability to establish and maintain effective working relationships with coworkers, owners,andclient. 14. Performsotherrelatedworkasassigned. JOBSPECIFICATION 1. Mayrequireahighschooldiplomaoritsequivalentwith02yearsofexperienceinthe fieldorinarelatedarea. 2. Abilitytounderstandandfolloworalandwritteninstructions. 3. Has knowledge of commonlyused concepts, practices, and procedures within a particularfield. 4. Reliesoninstructionsandpreestablishedguidelinestoperformthefunctionsofthejob. MESSENGER SUMMARY: Thisisworkinvolvingtheoperationofamotorvehicletotransportindividualsand/ora varietyofsupplies,products,ormaterials. DUTIESANDRESPONSIBILITIES: 1. 2. 3. 4. 5. 6. 7. 8. 9. ResponsibleforXrayfilm&ECGTesttobringtothephysicianforreading. Transportsstaffwithresponsibilityforassistingstaff. Responsibletobringanddeliverthedocumentsforappropriatepersons. Payingofelectricbill,CATV,WaterBill,TelephoneBill,PagIbig,SSS,Philhealth,BIRand othertasksassignedbytheofficemanager. Deliveringlaboratoryresultstoothermedicalcentreasrequested. Verifiesdescriptionandquantitiesofallitemspickedupordelivered. Maintains motor vehicle and related equipment by identifying, scheduling, and/or makingneededrepairs. Exercises independence in performance of responsibilities; receives administrative direction. Performsotherrelatedworkasassigned.

- 23 -

EZLabManual JOBSPECIFICATION: 1. Possession of a valid driver's license appropriate for vehicle assignment at time of appointment. 2. Oneormoreyearsofexperienceintheoperationofamotorvehicle;andcompletionof highschoollevel. 3. Comprehensiveknowledgeoftherulesandregulationsregardingthesafeoperationof motorizedequipment,andofthetrafficlawsoftheStateofPhilippines. 4. Abilitytooperateamotorvehiclesafelyandefficiently. 5. Abilitytoperformroutinemaintenanceonvehiclesandequipment. 6. Abilitytomaintainrecordsandpreparenecessaryreports. 7. Abilitytofollowsimpleoralandwritteninstructions; 8. Willingness to learn and perform tasks assigned; clerical aptitude; legible writing; alertness;neatappearance;tactandcourtesy;goodphysicalcondition. 9. Abilitytolift,moveandstackheavyobjects. 10. Abilitytoestablishandmaintaineffectiveworkingrelationshipswithothers. 11. Abilitytounderstandandcarryoutinstructions.

- 24 -

EZLabManual

COMPANYPOLICIES

- 25 -

EZLabManual POLICIESONPROBATIONARYEMPLOYEE

PROBATIONARYSTATUS

Theinitial(6)sixmonthprobationaryperiodforclassifiedstaffisservedonly onceduringcontinuousemployment. When a temporary employee moves to a regular position within the company, he/shemustcompleteasixmonthprobationaryperiodfromthedateofhis/her newregularposition. Ifanemployeetransfersorchangespositionswithinthecompanyduringhis/her sixmonth probationary period, he/she must complete the initial sixmonth probationaryperiod. An individual who leaves the company and is subsequently rehired must completeanewsixmonthprobationaryperiod. An employee may be terminated at the discretion of the company atany time during the employees probationary period. An employee serving the initial probationaryperiodisnotentitledtoappealorgrievancerightsintheeventof termination of employment. Further, a probationary employee does not have accesstothecompanysappealorgrievanceproceduresforotherissuesrelating toemployment. The superior is required to evaluate the employees performance during the probationary period and complete at least one performance appraisal form within30calendardayspriortotheendoftheprobationaryperiod. An employee who is reclassified in his/her current position is not subject to a newprobationaryperiod.

NEWEMPLOYEEPOLICIES

NEWEMPLOYEEBENEFITSORIENTATION All new benefitemployees shall attend an orientation session conducted by Human Resources Department. Contact the Human Resources Department for the date and time of thenextregularlyschedulednewemployeeorientation. SUPERVISORSORIENTATION The supervisor is responsible for the orientation and training of all new employees to thejob. GENERALPROCEDURES Thehiringsupervisormustensureeachnewemployeeperformstheappropriateinitial employmentprocedures. - 26 -

EZLabManual OPPORTUNITIESFORINTERNALCANDIDATES The company seeks to provide opportunities for all employees to be considered for promotion and/or advancement. The company believes in and is committed to a policy of upwardmobilityforitsinterestedemployeesthroughpromotionandtransferopportunities. JOBREFERENCES Supervisors may provide requesting employer information concerning a current of former employees reason for termination, job performance; professional conduct and evaluation are used for the purpose of evaluating the employee for employment. Any informationprovidesoncurrentorformeremployeesshallbejobrelated,truthful,factualand demonstrablefromtherecordsoftheemployee. APPLICANTS An applicant is defined as someone who has completed an EZ Lab Diagnostic & LaboratoryClinicemploymentapplicationandhassubmitteditinatimelymannerinresponse toanopenjobposting. EMPLOYMENTELIGIBILITY It is the companys responsibility to ensure that the identity and the employment eligibility of all persons employed by the company have been reviewed and that they are eligible for employment. Hiring officers shall be responsible for compliance with the documentationandverificationrequirementsofthispolicy. TRAINEE A managermay also choose to fill certain vacancies with a trainee. Atrainee may be hiredatarateofpaybelowtheminimumrateofpayforthatpositionforupto1monthwhile the employee is in training. A manager interested in filling a position with a trainee should informtheHumanResourcesDepartmentpriortocompletingthePersonnelRequisitionsothis may be considered in recruiting efforts. However, this decision may also be made after the applicantpoolhasbeenreviewed. GENERALPOLICY EZLabDiagnostic&LaboratoryClinicspolicyistocompensateemployeesonthebasis ofindividualperformance,market,applicablegovernmentalpolicies.

- 27 -

EZLabManual PAYRATECHANGES LegislatedPayRaises:certainsalarychangesoccurthroughstateappropriations.These may include, but are not limited to, general adjustments, merit pay adjustments and market pay adjustments. Eligibility for salary adjustments is based on legislation as well as company procedures. EZ Lab Diagnostic & Laboratory Clinic initiated pay raises: employee Salaries may be increased through a reclassification or a salary adjustment (a Salary within the same job classification).Reasonsforsuchadjustmentsmayinclude,butarenotlimitedto,assignmentof additionalresponsibilities(careerprogression),marketconsiderations,orequity.Forclassified staff and service professionals, the Human Resources Department shall determine an appropriatesalaryincreaseamount. Everyyear,allpermanentemployeesareentitledtoreceiveanadditionalP50increase onhis/hersalaryperday. ESTABLISHMENTOFPAYRATES NewHirePayRates:thehiringdepartmentwillestablishhiringpayrateswithinputfromthe Human Resources Department. Some considerations include: experience of the applicant, responsibilitiesoftheposition,applicablesalaryrange,marketandavailablefunding. Promotion:whenanemployeeispromoted,anappropriatepayratewillbeestablished,using thesameconsiderationasinestablishinganewhirepayrate. Trainee:atraineemaybehiredatapayratebelowtheminimumrateofpayornormalrateof payforthatpositionforupto1month. WORKINGPOLICIES Expectedworkhours: Therewillbe11hoursworkaccomplishmentper6daysaweek.7:00AMto6:00PM.Thismay bemodifiedbyamanagertomeettheneedsofthecompany. TheWorkweek The companys workweek consists of six days that begin on Monday and end on Saturdayfrom7:00AMto6:00PM.Thenormalworkscheduleforfulltimeemployeesconsists ofsixeighthourworkperiodsforatotalof48hoursworkedwithinthecompanysestablished workweek.Thismaybemodifiedbyasupervisortomeettheneedsofacompany.

- 28 -

EZLabManual Reportingtime/Requirements: 1. Thestaffisexpectedtobeattheclinicexactlyat7AMto6PM. 2. Signingoftheattendanceinlogbookshouldbedonepersonally. 3. Thestaffisrequiredtoreportinmajoruniform. RestPeriods An employee may have a compensable rest period of up to fifteen minutes each morning and fifteen minutes each afternoon during regularly assigned duty hours. The rest periodisintendedtobearecessfollowinganextendedperiodofworkandshouldbefollowed byanotherworkperiod.Restperiodsmaynotbeusedtocoverlatearrivalsorearlydepartures fromtheemployeesworkstationortoextendamealperiod.Restperiodarenotcumulative and,ifnottaken,theyareforfeited;theymaynotbecombinedtoprovideathirtyminuterest periodonceaday,normaybeaccumulatedforvacationorothertimeoff.Restperiodsmust betakenattheworksite.Worktimemaynotbeusedtotraveltoanotherareatotakethe fifteenminuterestperiod. MealPeriods Employeesareauthorizedtotakeaonehourunpaidmealperiodeachworkday. Emergencyleaves/absences/late: 1. Thestaffshouldbeinformedatonceincasethestaffwillgoonemergency leavetotheheadoftheoffice. 2. Allemployeesarerequiredtoreportabsencessuchasvacations,sick leave,andpersonaltimeoffontheLeaveApplicationForm(LAF) 3. RelieversareallowedonceMedicalTechnologistsareabsent. 4. Annualvacationleaveshouldbefiledahead3daysbeforetoallowtime forthestaff. 5. Prescribedformsickleavesshouldbeaccomplished. 6. Allformsforleaves/absencesshouldbedulysignedbytheheadofthe office. 7. Personnelcomingtoworklaterthan15minutesofdutyschedulewill receivesalarydeductionequivalenttoonehour.

- 29 -

EZLabManual Discount: 1. 20%discountofSeniorCitizens,presentationofI.D.isrequired 2. 100%discountofpartnersandstaff. 3. Immediaterelatives,familiesandfriendscouldavailtheir10%discountoflaboratory checkup. 4. Theallottedbudgetforoneyearhasacostsvalueof6,000.00.Ifitisalreadybeen consumedbythepartnersandstaff,theyareresponsibleforitspayment. RecordofAbsences A(LAF)mustalsobecompletedbyallemployeesforeachabsencefromtheworkplace. Wheneverpossible,allabsencesmustbeapproved,inadvance,bythemanagers&officerof themonth.Whennotpossible,absencesshouldbereportedassoonaspossible.Recordsof theseabsences,plushoursworkedbydate,mustbepostedtoanEZLabEmployeeTimeBook Record. TimeBookRecordRequirements Eachtwoweekpayperiod,mustcompleteandsignatimerecord.Managermustkeep recordsofcompensatorytimeearnedandtaken,aswellascurrentbalancesforeacheligible employee;theEZLabEmployeeTimeBookRecordmustbeusedtorecordhoursworked,hours paid,andovertimeearned.Theseformedwillbeexaminedinperiodicaudits,soitisimportant thattheybecompletedandfiledregularlytocomplywiththeFairLaborStandardsAct. GENERALINFORMATION EZLabDiagnostic&LaboratoryClinicofferseligibleemployeesavarietyofbenefitplans. Tobeeligibleforbenefits,anemployeemustbearegularemployeeworking40hoursormore per week for six months or more. These benefit programs are available to all company employees. ContinuationofBenefits EZLabDiagnostic&LaboratoryClinicoffersfreemedicallaboratoryandcheckuptoall employeesamountingtotwelvethousandpesos(P12,000.00)foroneyear.Thecompanyalso offers10%discounttoallrelativesoftheemployee. RequiredBenefits The company is required by law to provide certain benefits for employees. Social Security,PhilhealthInsuranceandPagIbigarecostpaidbycompanyandemployeeequally.

- 30 -

EZLabManual HOLIDAYS OfficialcompanyholidayshavebeendesignatedbythePresidentasfollows: RegularHolidays NewYearsDayJanuary1 MaundyThursdayMovableDate GoodFridayMovableDate EidulFitrMovableDate ArawngKagitingan(Bataan&CorregidorDay)MondaynearestApril9 LaborDayMondaynearestMay1 IndependenceDayMondaynearestJune12 NationalHeroesDayLastMondayofAugust BonifacioDayMondaynearesttoNovember3 ChristmasDayDecember25 RizalDayMondaynearesttoDecember30 SpecialHolidays NinoyAquinoDayMondaynearesttoAugust21 AllSaintsDayNovember1 LastDayoftheYearDecember31 In the event the holidays fall on a Wednesday, the holiday will be observed on the Monday of that week. If the holiday falls on a Sunday, the holiday will be observed on the Mondaythatfollows. The company will officially be closed for business during the Regular Holidays, and employees will receive pay for these days off. The business will be opened during Special Holidays. EligibilityforHolidayPay Allregularemployeesareeligibleforholidaypayprovidedtheyareinpaystatusforthework dayprecedingandtheworkdayfollowingtheholiday.Employeesonsickleaveareconsidered to be in pay status for this purpose. All unconsumed sick leave in a year are convertible to theirregulardayratemultipliedbyfive(5)daysonly. HolidayduringPaidLeave IfarecognizedCompanyholidayfallsonaregularworkdayduringpaidsickleaveperiod,the dayispaidasaholiday,andisnotcountedagainstpaidleavebalances. HolidayduringUnpaidLeave IfarecognizedCompanyholidayfallswithinanauthorizedleavewithoutpayperiodthatdoes notexceedthedurationofthenormalpayperiod,anemployeeispaidfortheholiday. - 31 -

EZLabManual If a recognized Company holiday falls on a Monday on which an employee is scheduled to returntoworkfromanauthorizedleavewithoutpayofanyduration,andtheemployeereturns toworkontheholidayoronhisorhernextscheduledworkday,theemployeeispaidforthe holiday. HolidayPayforTerminatingSupportStaffEmployees IfarecognizedInstituteholidayfallsonthelastavailablescheduledworkdayofapayperiod, and the employee is terminating on his or her preceding work day, he or she is paid for the holiday. HOLIDAYPAYPOLICIES HolidayPay StaffmembersarepaidforrecognizedCompanyholidaysaspartoftheirmonthlysalary,when theyareinpaystatus. HolidayduringVacation If a recognized Company holiday falls during a staff member's vacation, the day is paid as a holiday,andisnotcountedagainstvacationallowances. HolidayduringUnpaidLeave If a recognized Company holiday falls during an authorized leave without pay period, a staff memberisnotpaidfortheholiday. IfarecognizedCompanyholidayfallsonthedayastaffmemberisscheduledtoreturntowork fromanauthorizedleavewithoutpayofanyduration,andthestaffmemberreturnstoworkon theholidayoronhisorhernextscheduledworkday,heorsheispaidfortheholiday. HolidayPayforTerminatingStaffMembers IfarecognizedInstituteholidayfallsonthelastavailablescheduledworkdayofthemonth,and thestaffmemberisterminatingonhisorherprecedingworkday,heorsheispaidtotheend ofthemonthtoincludetheholiday. VACATION Fulltime employees are eligible for 15 days of vacation leave for each year. Part time employeesarenoteligibletovacationleave.Employeesareresponsibletosubmitleaveform prior to the date of vacation leave. Managers have responsibility for coordinating and approvingvacationschedulesoftheiremployees. HolidayorIllnessduringVacation If a recognized company holiday falls on an employee's regularly scheduled workday during his or her vacation, the day is paid as a holiday and is not counted against vacation - 32 -

EZLabManual balances. Ordinarily, illness during vacation is not counted as sick leave. However, if an employeehasaseriousaccidentorillnesswhichimpairstheintendedbenefitsofthevacation, themanagershoulddiscussthematterwiththeManagementCommitteetoconsiderwhether sickleaveshouldbegranted,andpartorallofthevacationrescheduled. Asmentioned,theemployeeisresponsibleforcheckingthenumberofvacationdayand thenseekingpermissionfromthemanagerforvacation.Theemployeeisalsoresponsiblefor informingthemanagerwhenheorsheistakingdaysofvacationcredit.Generally,themanager isresponsibleforrecordingthenumberofapprovedvacationdaystaken. SICK/PERSONALLEAVE Thecompanygrantspaidsickleavetoemployeestoguardagainstlossofearningsdue toillness.Managershavetheprincipalresponsibilityfortheproperandconsistentapplication ofthesickleavepolicy.Abuseofsickleaveinanyformcanhaveseriousimpactonthemorale andeffectivenessofallmembersofagrouporoffice. NotificationofAbsence Anemployeemustnotifyhisorhermanageronorbeforethefirstdayofabsencedue to illness in order to be paid for sick leave. In cases of prolonged illness, employees are expected to keep their managers informed at regular intervals of their condition and their anticipateddateofreturntowork,sothatworkmaybeproperlyscheduledorreassigned. PaidLeaveforMedicalExaminationandCare It may be necessary from time to time for an employee to seek medical care during workinghours.Thecircumstancesofeachincidentdeterminewhetherthetimeawayfromthe jobshouldbepaidastimeworkedorshouldbechargedtosickleavebalances. MedicalCertification Themanagerisresponsiblefortheequitableadministrationofthecompany'ssickleave policy, and may require medical certification from an employee's (or family members) physicianindicatingthattheemployeeisunabletoworkbecauseoftheemployeesorfamily membersillness,beforethesickleaveispaid. IllnessduringVacation Ordinarily,illnessduringvacationisnotcountedassickleave.However,ifanyemployee hasaseriousaccidentorextensiveillnesswhichdestroystheintendedbenefitofthevacation, themanagershoulddiscussthematterwiththeManagementCommitteetoconsiderwhether sickleaveshouldbegranted,andpartorallofthevacationrescheduled. SickLeaveforDisabilityAssociatedwithPregnancyorChildbirth Disabilitiesassociatedwithchildbirthwillbetreatedasanyotherdisabilitywithregard topaymentforsickleaveinaccordancewithapplicablesickleavepolicies. - 33 -

EZLabManual AmountofSickLeave Fulltime and parttime employees will be credited with 15 days of sick leave on the employee's anniversary date of each year, provided that they have completed at least six monthsofserviceattheCompany.Sickleaveispaidatnormalratesandmustbereportedon attendancerecords. UseofSickLeave Sickleaveistobeusedincasesofaccidentorillnessoftheemployee,andmedicalor dentalexaminationandcare.Inaddition,anemployeemayuseuptofifteendaysofsickleave everyyeartocareforasickfamilymember,suchasachild,spouse,parent,parentinlaw,or othermemberofthehousehold,andfortheirmedicalordentalexaminationandcare. HolidaywithinSickLeave Ifarecognizedcompanyholidayfallsonaregularlyscheduledworkdayduringapaid sickleaveperiod,thedayispaidasaholiday,andisnotcountedagainstsickleaveallowances. PERSONALLEAVEPOLICY PersonalLeaveforSupportStaff EZLabDiagnostic&LaboratoryClinicspolicyonpersonalleaveisintendedtoprovide Staffwithflexibilityintheirworksituationsbyallowingtoaccommodatepersonalneedswhich otherwisewouldhavetobechargedtovacationbalances. ONABSENCESANDLEAVE NoticeofAbsence Inkeepingwithotherleavepolicies,Staffwillberequiredtoobtaintheapprovaloftheir managers to be absent on personal leave on a given day or part of a day. It will normally be expectedthatemployeeswillobtainthisapprovalatleastthreeworkingdaysinadvanceofthe plannedabsence.Noexplanationofthereasonsfortheabsencewillberequiredexceptwhen emergency situations cause employees to seek approval for absences on shorter notice. Personalleavemaybetakeninlessthanfulldayperiodsandshouldbereportedtothenearest quarterofanhour. Leave Related to an Employee's Own Serious Health Condition (Including Pregnancy and Childbirth) Inthecaseofanemployee'sownserioushealthcondition,includingconditionsrelated to pregnancy and childbirth, leave may be approved in continuous periods, intermittent periods,orbyareducedworkschedule,inaccordancewithcertificationfromtheemployee's health care provider. In cases of reduced schedules or intermittent leave, a manager may transfertheemployeetemporarilytoanalternatepositiontoaccommodatetheneedsofthe - 34 -

EZLabManual workarea.Leaveswhichextendbeyond12weeksindurationduetoanongoingserioushealth condition will be reviewed on a casebycase basis in accordance with EZ Labs general sick leavepoliciesasoutlinedonthisManual. Sickleavepoliciesfortheparticularpayrollcategoryshouldbereviewedindetermining anypaidleaveforwhichtheemployeeiseligible.Ifanemployeeisnoteligibleforpaidleave,or ifpaidsickleavebenefitshavebeenexhausted,theleaveperiodwillbeadministeredasunpaid leave. Followingamedicalleaveperiodrelatedtopregnancyandchildbirth,anemployeemay be eligible for additional leave, without pay, for purposes of child care. In such cases, the combinationofthemedicalleaveperiodandtheadditionalperiodforchildcareleaveshould notexceed12weeksintotal. LeavetoCarefortheEmployee'sNewbornChild Leave to care for a newborn is unpaid leave. Following a sick leave period related to pregnancy and childbirth, an employee may be eligible for additional leave, without pay, for purposes of child care. In such cases, the combination of the medical leave period and the additionalperiodforchildcareleaveshouldnotexceed12weeksintotal.Requestsforleaveto exceed12weekswillbereviewedinaccordancewiththespecificcircumstancesoftherequest, considerationsofworkcoverage,andtheprovisionsoftheCompany'sgeneralpolicyonleave withoutpay. LeavetoCareforaFamilyMemberwithaSeriousHealthCondition Employeesmayrequestleaveinsituationswheretheyarerequiredtoparticipateinthe careofaseriouslyillchild,parentorspouse.Requestsforleavetocareforotherseriouslyill familymembersormembersofanemployee'shouseholdmaybeconsideredonacasebycase basis. An employee may request leave in continuous periods, intermittent periods, or by a reducedworkscheduletoatotalof12weeks,inaccordancewithcertificationfromthefamily member's health care provider. In cases of reduced schedules or intermittent leave, a department supervisor may transfer the employee temporarily to an alternate position to accommodatetheneedsoftheworkarea. Leave periods to care for seriously ill family members are unpaid leaves. Requests for leavetoexceed12weekswillbereviewedinaccordancewiththespecificcircumstancesofthe request,considerationsofworkcoverage,andtheprovisionsoftheCompany'sgeneralpolicy onleavewithoutpay RequestandAuthorizationofEZLabLeave Employeesareexpectedtoprovideasmuchadvancenoticeoftheneedforleaveasis possible to allow for planning of coverage in the work area. Notice of at least two weeks is - 35 -

EZLabManual preferred.LeaveofAbsenceAuthorizationformsareavailableintheOffice. In addition, certification from a qualified health care provider will be required for approvalofaleavetocareforaseriouslyillfamilymemberorinthecaseofanemployee'sown serious health condition. Such certification should be provided prior to the beginning of the leave (or in as timely a manner as possible) and should include: the date on which the illness/conditioncommenced;theestimateddurationoftheillness/condition;astatementthat theillness/conditionwarrantstheparticipationoftheemployeetoprovidecare(inthecaseof an ill family member); and, in cases in which intermittent leave is advised, the dates and durationoftreatmentstobegiven.CopiesoftheCertificationofHealthCareProviderformare availableforthispurpose. ONACCIDENT&COMPENSATION IndustrialAccidentandWorker'sCompensation The company provides insurance to protect employees against financial loss due to personalinjuryandsomeoccupationalillnessesarisingoutofandinthecourseofemployment attheCompany. ReportingaLaboratoryAccident Anemployeewhohasbeeninjuredorbecomesillasaresultofhisorheremployment at the Company should notify his or her manager at once, and seek immediate medical attention.Themanagermustalsosubmitawrittenreportoftheillnessorinjurythatoccurred. BEREAVEMENTLEAVE Eligibility Bereavementleavepolicyisdesignedtoprovideemployeeswithpaidtimeawayfrom worktogrieveandtohandlemattersrelatedtoadeathintheirfamily.Allfulltimeemployees areeligibleforbereavementleave. LengthofBereavementLeave In the event of a death in an employee's family, after discussion with his or her manager,theemployeemaybegranteduptofivedaysofpaidleaveforbereavement. A day of bereavement leave is based on the standard workday the employee would have workedontheday(s)takenforbereavement. PaymentofHolidayPay Anemployeewillbepaidforaholidaywhichfallsduringaperiodofabsenceduetoan occupational injury or illness only if he or she is supplementing his or her wage replacement

- 36 -

EZLabManual benefitswithsickleave.Inthisevent,theportionofsupplementalpaynormallychargedtosick leavewillbechargedtoholidaypay. ReviewofEmploymentStatus Incasesofprolongedabsenceduetoanoccupationalillnessorinjury,theemployment status of the individual will be reviewed periodically, and may result in termination of employmentwiththeCompany. LEAVEOFABSENCEWITHOUTPAY ApprovalofLeavewithoutPay Leavesofabsencewithoutpaymaybegrantedforaspecifiedperiodoftime,normally for educational or public service reasons and because the Company wishes to maintain an associationwiththeindividual.Amanagermayauthorizeanindividualtotake,inanycalendar year,unpaidleaveofdurationnottoexceedtheindividual'snormalpayperiod.Requestsfor leavesmustbeapprovedbythemanagers. Eligiblestaffmembersareentitledtoaleaveofabsencewithoutpayofupto12weeks in a 12month period to care for their newborn child or a child newly placed with them for adoptionorfostercare;becauseoftheserioushealthconditionoftheirchild,parent,orspouse requiringtheirparticipationincare;orbecauseoftheirownserioushealthcondition,including conditionsrelatedtopregnancyandchildbirth. RequestforLeavewithoutPay A request form for Leave of Absence Authorization, available from the Office of PersonnelServicesshouldbecompletedatleasttwoweekspriortotheanticipatedbeginning ofaleavewithoutpay.Allsignaturesnecessaryforauthorizationoftheleavemustbeobtained by the employee before the form is forwarded to the Office of Personnel Services for final approvalandprocessing.Theformcontainsspecificinstructionsforitscompletion. One copy of the signed form is returned to the department in which the employee works, one copy is returned to the employee, and one becomes part of the employee's personnelfileintheOffice. PAYMENT AneligibleemployeewhoworkedduringSpecialHolidaywillbepaidattherate of130%ofhis/hersalary. Holidaysfallingwithinavacationorsickleaveperiodwillbetreatedadholidays; therefore,theywillnotbechargedtovacationorsickleave. Upontermination,anemployeewillbepaidanyholidaypaythathe/shehadnot receivedandforwhichhe/shewaseligible. - 37 -

EZLabManual AccommodationofReligiousObservances/Practices No employee shall discriminate against any employee or other individual because of suchindividualsreligiousbelieforpracticeoranyabsencethereof.Furthermore,amanageris responsibletoreasonablyaccommodateindividualreligiouspractices. ELIGIBILITY Regularemployeesworking40hoursperweekormoreforacontinuousperiod of six months or more are eligible to use vacation. However, employee is not eligibleforvacationpay. CarryForwardofAccruedVacation An employee is encouraged to use his/her vacation hours each year, rather than permitting vacation to accumulate. Unused vacation leave may be accumulated and carried forwardfromonecalendaryeartothenext. TimeCharges AllemployeesarerequiredtocompleteaRecordofAbsence(ROA)formforall vacationandsickleavetaken. Sickleavewhileonvacationisauthorizedonlywhentheemployeehasamajor illness or injury and is unable to continue the vacation activity. A physicians writtenstatementmayberequiredtoverifysuchanillness. Holidaysfallingwithinavacationperiodwillbetreatedasholidaysandtherefore notbechargedtovacationleave. ONVACATION SCHEDULINGOFVACATIONTIME Anemployeemustrequestapprovaltousevacationtimeinadvanceoftakingtimeoff. Scheduling of vacation time will be the responsibility of the manager concerned. While vacation will be granted, whenever possible, to satisfy individual requests, continuity of operations must be maintained. Consequently, vacations must be scheduled in accordance withtherequirementsofthecompanyfortheemployeesservices. REPORTSANDRECORDS Managers are responsible for recording and maintaining accurate records of vacation taken by their employees. This responsibility includes timely reporting of hours taken and monitoringaccruals.Acompanytimekeepermaybeassignedtoassistamanagerinthistask butitremainshis/herresponsibility. - 38 -

EZLabManual ELIGIBILITY

Regularemployeesworking40hoursperweekormoreforacontinuousperiod ofsixmonthsormoreareeligibletoaccrueandusesickleave. Temporaryemployeesarenoteligibleforpaidsickleave.

CONDITIONSOFUSAGE Sick leave benefits are designed to provide for continuation of income during times of legitimate illness. Sick leave is not to be considered as an alternative formofvacationtime,holidayleave,ortimeoffpersonalbusiness. An eligible employee may be granted sick leave when he/she is unable to perform his/her duties because of illness, injury, pregnancy, childbirth, or becausehe/shemustbeabsentfromworkforthepurposeofobtaininghealth relatedservicesnotavailableafterregularworkinghours. Sick leave may be granted to an eligible employee whose absence is due to illness or communicable disease within the employees immediate family or household. Immediate family is defined as a parent, parentinlaw, brother, sister, spouse, child, grandparent, grandchild, and any other person who is a permanentmemberoftheemployeesestablishedhousehold. Sickleaveshallnotbeallowedforabsenceduetoanyofthefollowing: 1. Disabilityarisingfromanysicknessorinjurypurposelyselfinflicted orcausedbytheemployeeswillfulmisconduct. 2. Sickness or disability while on layoff status, leave of absence withoutpay,orvacation,exceptwhentheemployeehasamajor illnessorinjuryandisunabletocontinuethevacationactivity.A physicians written statement may be required to verify such an illness. 3. Sick leave should be requested and approved in advance, when possible. Sudden illnesses that result in absences must be reported to the employees manager within three working days after an employees return from illness. In situation where an employeeisunabletocompleteaRecordofAbsenceformtomeet biweeklydeadlines,themanagermaycompletetheformforthe employeetoensurecontinuityofpay. 4. For absence in excessof three days becauseof illness, injury, or other disability, or should there be reason to believe that sick leaves is being abused, the employee may be required to furnish thecompanywithawrittenstatementfromaphysicianmustalso certifythattheemployeeisabletoreturntowork.Intheevent the manager exercises this option, the employee will not be permittedtoreturntoworkwithoutthisphysiciansstatement. - 39 -

EZLabManual REPORTINGANDRECORDKEEPING ItistheemployeesresponsibilitytosubmitanaccurateRecordofAbsenceform reportingthehourshe/shedidnotworkforwhichsickleaveisrequested. Itisthemanagersresponsibilitytorecordandmaintainaccuraterecordsofsick leavetakenbyhis/heremployees.Thisresponsibilityincludestimelyreporting of hours taken and monitoring accruals. A department timekeeper may be assigned to assist a manager in this task but it remains the managers responsibility. PAYMENT Paymentforsickleaveapprovedandtakenismadeattheemployeesstraight timerateofpayfortimelostwithintheregularlyscheduledworkperiod. Ifaholidayfallswithinaperiodofapprovedsickleave,itischargedasaholiday, notasickleavetime. RETURNTOWORK The employee will be returned to his/her former or essentially equal position uponreturningfromleave.Iftheleaveisbasedonmedicalneeds,theemployee isresponsibleforprovidingadequatedocumentationtosupportreturntowork and/orfitnessforduty. If a company head has the reason to question the capacity of an employee to perform duties and responsibilities, the company head may choose to require the employee to be examined by a physician of the companys choosing for a fitnessfordutyexamination. FAILURETORETURNORACCEPTEMPLOYMENT Anemployeewillbeconsideredtohavevoluntarilyterminatedemploymentifhe/she: a) FailstoreturntoworkupontheexpirationoftheExtendedLeaveofAbsenceordate agreedupon,;or b) Failstocontracthis/hercompanyheadtwoweekspriortotheendoftheExtended LeaveofAbsencetorequestanextension(ifapplicable);or c) Advisesthecompanyofhis/herintentionnottoreturntowork.

- 40 -

EZLabManual ABSENCEWITHOUTPAY Absencewithoutpayisusefortemporaryabsencesonlyandmaybeusedforuptoone payperiodsworthoftimeforoneevent. Requestsforabsencewithoutpaymustbemadeinadvanceandmusthavethe approvalofthemanagerandthecompanyhead. EMPLOYEEPERSONNELRECORDSMANAGEMENT OFFICIALPERSONNELRECORDS The Human Resources Department maintains an official personnel record for probationaryandregularemployees. CONTENTSOFOFFICIALANDOTHERPERSONNELRECORDS Typically,eachemployeesofficialpersonnelrecordcontainsthefollowing: 1. All job application forms, resumes, vitae, transcripts, letters of reference and otheremploymentrelateddocumentssubmittedtothecompany. 2. Copies of all performance appraisal reports for classified staff and service professionals. 3. A chronological file, record, or copies of approved personnel action forms, memoranda,orotherdocumentsthatreflectauthorizedchangesinemployment status,salary,etc.;whensuchdocumentsareavailable. 4. Official correspondence with or about the employee, including any letters of recommendation, reprimand, or employee comments concerning statements containedinthefile. 5. Documents acknowledging or verifying receipt by an employee of disciplinary communications. (Warning, either verbal or written, is not considered disciplinarycommunications.) 6. Documents, certificates, diplomas, etc., related to further education, trainings andcertificationcompletedbytheemployee. 7. Any other document, form, letter, memorandum, agreement, etc., that is deemed by the Director of Human Resources to be an employee record that shouldbeplacedintheofficialpersonnelrecord. PROCEDUREFORVIEWINGEMPLOYEEFILES If an employee wishes to view the contents of his/her Human Resources Departmentfile,he/sheshouldsetupanappointmenttodosowitharepresentativeof the Human Resources Department. The employee should not assume that the staff memberwillmakecopiesofanyitemsinthefileduringthisviewing,butmayrequest specificcopiestobepickedupatalaterdate.Achargemaybemadeforcopies.

- 41 -

EZLabManual PERSONNELSTATUSCHANGE An individual employee and his/her supervisors should keep the Human ResourcesDepartmentinformedofanychangesinpersonalstatus(i.e.;marital, numberofdependents,etc.).Anybirthsordeathsinanemployeesfamilyora changeinmaritalstatusmayaffecttheemployeesincometaxwithholding.An employee may add dependents to his/her life insurance policies; if the Human Resources Department is notified within 30 days of the date such dependents first become eligible. Designated beneficiary changes can be made at anytime andshouldbereportedtotheHumanResourcesDepartment. A change in name, address, telephone number, emergency contact name, and/or address should be reported to the Human Resources Department immediately.Itisimportantinemergencysituationsthatcurrentinformationbe available. Contact the Human Resources Department for assistance and clarification on benefits. INQUIRIESREGARDINGCURRENTORPASTEMPLOYEES TheHumanResourcesDepartmentistheofficialsourceofinformationregarding presentorformeremployees. Employment verifications requested by banks, or other financial institutions shouldbereferredtothePayrollOfficer. A copy or written communications regarding verification of past employment must be sent to the former employees last known address. This will include boththeinformationrequestedandsupplied. EMPLOYEEDEVELOPMENTANDTRAININGPROGRAM The company is committed to employee professional development. In an effort to accomplish this, the Human Resources Department sponsor a broad range of program opportunitiesincludingworkshops,seminars,classesandcomprehensivecoursesofstudy. MANDATORYTRAINING Managers are required to allow classified staff and service professional employeestoparticipateincareerandprofessionaldevelopmentprogramseach fiscal year and to support developmental leaves for those employees participating in these programs. Time taken to attend workshop must be approved in advance by the managers. While all managers are encouraged to allowemployeesofffromworktoenhanceprofessionaldevelopment,continuity of operations must be maintained. With prior manager approval, employees shall be allowed to use work time rather than take vacation time or leave withoutpaytoattendtrainingprogramsrelatedtotheirjob.

- 42 -

EZLabManual Government legislation mandates that company heads ensure all employees (regular, temporary, and student workers) receive Public Service Orientation (Ethics) training at least once every two years. Employees must attend Safe WorkingandLearningEnvironmenttrainingatthebeginningofemployment. RECOMMENDEDTRAINING Managersshouldparticipateinprogramsregardingculturaldiversity,affirmativeaction, performance appraisals, and general management/supervisory issues to gain perspective on thecompanyspoliciesandphilosophiesintheseimportantareas.Managersareencouragedto contacttheHumanResourcesDepartmentformoreinformation. PERFORMANCEREVIEWGUIDELINES Inworkperformancediscussions,thefollowingtopicsareusefulforbothmanagersand employeesandshouldbethebasisofadiscussionatonceamonth.Ideally,communicationand feedbackexistcontinuouslybetweenofficerofthemonthandemployee.However,evenwhen feedback is freely given and received, a more formal annual or semiannual performance reviewisdesirable.Itisthemanager'sresponsibilitytoseetoitthatperformancereviewstake placewithallemployees. It is important for both managers and employees to have considered the following elements priortotheactualperformancereviewdiscussionandbepreparedtodiscussthem. CLASSIFIEDSTAFFPERFORMANCEAPPRAISALS APPRAISALSCHEDULE The performance of each regular employee will be appraised at least once a year. Classified staff appraisals are due to the Human Resources Department betweenNovember1andJanuary31.TheHumanResourcesDepartmentstaff may notify the appropriate administrative levels of delinquent performance appraisals. An employee may be evaluated more frequently at the managers discretion,orattheemployeesrequest. Anytime an employees performance changes significantly, a new performance appraisal from should be completed and submitted to the Human Resources Department. PROBATIONARYPERFORMANCEAPPRAISALS All newly hired classified staff employees will receive a probationary performance appraisal due within 30 calendar days prior to completing six months of continuous service. Aftertheprobationaryperiodiscompleted,allclassifiedstaffwillreceiveanannualappraisal

- 43 -

EZLabManual duetoHumanResourcesbyNovember30ofeachyear.Aprobationaryemployeewillservea minimumsixmonthprobationaryperiod. Ifamanagerpermitstheemployeetoworkbeyondthesixmonthprobationaryperiod withoutaformalappraisaltotheHumanResourcesDepartmentthatemployeemayrequestan appraisal but shall not be moved to a regular status automatically. Employees transferring betweenbranchorpositionswithoutabreakinserviceshallnotserveasecondprobationary period. EMPLOYERRESPONSIBILITIES Thecompanyisresponsibleforprovidinganenvironmentreasonablefreefromhazards that may cause illness, injury, or death to employees, patients and visitors. As such, the Companywill: b. Establish and maintain safety and loss prevention/risk management programs that seek to protect human health, to ensure regulatory compliance and to preservecompanyinterestandassets. c. Incorporate educational programs and training designed to prevent accidents and injuries, monitor procedures to identify hazardous situations, and take correctiveactionstoeliminateknowndeficiencies. d. Comply with applicable Federal, State, Country, and local safety laws and regulations. e. Maintain an incident management plan that seeks to reduce risk to the health andsafetyofthecompanycommunityintheeventofanemergency. EMPLOYEERESPONSIBILITIES All employees are responsible for maintaining a safe environment through their work andotheractions.Assuch,employeesarerequiredto: 1. Complywithapplicablegovernmentallawsandregulations. 2. Observeallcompanyrulesandregulationsrelatedtotheefficient andsafeperformanceoftheirwork. 3. Attendassignedtrainingtopromotesafetyandminimizerisks. 4. Reporttomanagerand/orcorrecthazards,unsafeequipment,and improperworkpractices,asappropriate. 5. Reporttomanageraccidents/injuriesthatoccurtothemselvesor otherswhileonthejoboroncompanyproperty. 6. Report to police office and crimes and/or acts of violence or threatsofviolence. 7. Operatecompanyorpersonalvehicleswhileoncompanybusiness in accordance with transportation guidelines and applicable state andlocallaws. - 44 -

EZLabManual MANAGERRESPONSIBILITIES Allcompanyemployeeswhosuperviseothersareresponsibleformaintaininga safeworkenvironmentbyensuringthat: 1. Employees are trained in basic safety measures, fire and fire hazard prevention,andemergencyevacuationprocedures.Managersshouldensure thatjobspecificsafetytrainingisidentifiedandassignedtoemployees.This trainingmayincludeproperhandlingofchemicalsandbiologicalagents,and othertypesoftraining. 2. Safetytrainingrecordsaremaintainedfortheiremployees. 3. A manager report of injury is filed when an accident or workrelated health problemorinjuryoccursintheirareaofresponsibility. 4. Equipmentandpropertywithintheareaofresponsibilityaremaintainedina safecondition. 5. Employeesarerequiredtofollowsafeworkpracticesatalltimes. 6. If an employee violates safe work practices or procedures, the unsafe behavior is halted, the employee is informed of the infraction(s), and the correctmethodofworkisreinforced.Disciplineforfailuretoperformworkin asafemannerwilldependontheseverityoftheincidentandwillrangefrom verbal reinforcement of safe working procedures, up to and including termination. Smokingisdefinedasthecarryingoruseoflightedpipes,cigars,cigarettes,snuff,ortobaccoin anyform. POLICY 1. SmokingisprohibitedinEZLabbuilding. 2. Smokingisprohibitedinsideallcompanyownedvehicles. 3. Complaints or concerns regarding violations of this policy or disputes regarding its implementationshouldbereferredtothemanagerforresolution. 4. Violatorsofthispolicymaybesubjecttodisciplinaryaction. ALCOHOL/DRUGPOLICY 1. Any violation of the drug/alcohol policy can result in disciplinary action up to and includingtermination. 2. EZLabDiagnostic&LaboratoryClinicrequiresthatcompanyemployeesreporttowork in a condition to perform their duties safely and efficiently in the interest of their co workersandpatients,aswellasthemselves.Forthesereasons,theuseofillegaldrugs andalcoholisprohibitedduringworkhoursatthecompanyworkplace,whileoccupying companyvehicles,whileoncompanybusinessandduringworkingtime.

- 45 -

EZLabManual 3. All state, federal, and local laws and company regulations pertaining to alcohol and other drugs apply to all members of the company community including employees, partnersandguests. ALCOHOLPOLICY 1. Alcohol beverages may not be sold or consumed on property owned or controlled by the EZ Lab System except when permitted by the general manager of the respective companyordesigneeincompliancewithapplicablelaws. 2. Companyemployeeorguestwillberesponsiblefortheregulationsfoundinthepolicy manual. 3. No person may consume or be under the influence of alcohol while attending and fulfilling employment responsibilities. Employees must conduct themselves in a responsibleandprofessionalmanneratalltimes. DRUGPOLICY 24. Possession, use, sale, manufacture, transfer or cultivation of any types of controlled substances (to include, but not limited to, barbiturates, opiates, marijuana,amphetamines,orhallucinogens)oraidingintheuseofsuch,violates thelawsofthePhilippines. 25. Use of illegal drugs or the use of prescription medication or over the counter drugsisdangerousandunacceptablebehaviorinthecompanycommunityandis prohibited. 26. Useorpossessionofanycontrolledsubstanceorillegaldrugorparaphernaliafor illegaldruguse,andtheunauthorizeddistributionorpossessionforthepurpose ofdistributionofanycontrolledsubstanceorillegaldrugisprohibited. 27. Company employees must report to the General Managers any criminal drug statute conviction for a violation occurring in the workplace no later than five workingdaysaftersuchconviction. REPORTINGVIOLATIONSOFTHEALCOHOL/DRUGPOLICY Complaints made by company employees and/or guests about a company employee shouldbereportedtotheemployeesimmediatemanager. EMPLOYEESSUBJECTTOTESTING Anycompanyemployee(regularfulltimeorparttime,temporaryorstudentemployee whose job description involves a safetysensitive function or requires possession of a commercialdriverslicenseforemploymentpurposes,issubjecttodrugand/oralcoholtesting.

- 46 -

EZLabManual SUBSTANCETOBETESTED Substancestobetestedforunderthispolicyare: 1. Breathalcoholconcentration 2. Controlled substances (to include but not limited to) cocaine, marijuana, opiates (includingheroin)amphetamines,andphencyclidine. Inaccordancewithgovernmentregulations: 1. Noemployeeshallbeondutywhilepossessingorconsumingalcohol. 2. Noemployeeshallbeondutywhilepossessing,using,orexhibitinganytraceof controlledsubstancesinthebody. 3. Noemployeeshallreportfordutyorremainondutyrequiringtheperformance ofsafetysensitivefunctionswhilehavingabreathalcoholconcentration.Safely sensitive functions are defined as those which could endanger the life of the employee,otheremployees,patients,ormembersofthepublic. 4. No employee shall report to duty within a minimum of four hours after consumingalcohol. ALCOHOLANDCONTROLLEDSUBSTANCETESTINGREQUIRED 1. Preemployment Testing shall be administered after an offer to hirehas been made and prior to an employee actually performing safety sensitive functions for the first time. Testing is also required for employees who are transferred/promoted into a classificationrequiringacommercialdriverslicense.Allapplicantswhorefuseorfailto passapreemploymentdrugandalcoholtestwillresultintheemploymentofferbeing withdrawn. 2. PostaccidentTestingshallbeadministeredafteranaccidenthasoccurredwherethe employeereceivedamovingtrafficcitationandaftereveryaccidentinvolvingafatality, regardlessofwhowasatfault.Testingshalloccurwithinaminimumoftwohours,but nottoexceedaneighthourperiodfollowingtheaccident. 3. Reasonable Suspicion Testing shall be administered when a manager has a reasonablesuspicionthatemployee(s)areinviolationofthealcohol/drugpolicy,the employee(s)willbereferredtothetestingsiteimmediately. 4. AlltestingarecoordinatedbytheGeneralManagers. CONSEQUENCESOFTESTINGREFUSAL 1. NewHireswhorefusepreemploymenttesting,orfailtoappearfortestingwillhave the job offer withdrawn unless valid medical documentation is presented to the managerinadvanceornolaterthanoneworkingdayfollowingthescheduledtest. 2. Employeeswhorefusetesting,unlessvalidmedicaldocumentationispresentedtothe manager,willreceivedisciplinaryactionuptoandincludingtermination. - 47 -

EZLabManual DEFINITIONOFFITNESSFORDUTYEVALUATION Anevaluationmadebyaphysician,psychologist,orotherevaluatortodeterminewhetheran employeeisabletocontinueintheperformanceofallessentialfunctionsofhis/herjobwithor withoutreasonableaccommodation. GENERALGUIDELINES 1. Each employee is responsible for maintaining his/her health in such a way that the employeecanperformtheessentialfunctionsofhis/herjobwithorwithoutreasonable accommodation. If a manager has reason to question the capacity of an employee to performdutiesandresponsibilities,aFitnessforDutyEvaluationmayberequired. 2. To determine the appropriateness of a formal Fitness for Duty request, the manager mustconsultwiththecompanyphysicians. 3. If assistance is needed in working through manager and employee relationship problems,orifthereareconcernsabouttheemotionalorbehavioralimpactofaFitness for Duty Evaluation requirement on an employee, or more immediate concerns about the employees emotional/behavioral condition, the General Managers may be consulted. 4. IfthecompanyrequiresaFitnessforDutyEvaluation,theemployeewillberequiredto beexaminedbyanevaluatorofthecompanyschoosingatapredeterminedtimeand location. The company will pay the full cost of the examination. Failure to attend a Fitness for Duty Evaluation may lead to disciplinary action, up to, and including, termination. 5. Records,reportsandresultsofsuchanEvaluationwillbemaintainedconfidentiallyand separatelyfromtheemployeesofficialpersonnelrecord.Thereportisandwillremain companyproperty. 6. Time required by the employee to complete the Fitness for Duty Evaluation is consideredascompanyworktimeandmayrequireanadministrativeleave.Timeofffor prescribedtreatment(aftertheEvaluation),mandatoryorotherwise,willbechargedto accruedsickleave. 7. For absences in excess of 2 weeks the employee may be required to be on a leave status.The purpose of this policy is to cooperate with state and local officials in the lawfulcollectionofgarnishmentsandresolutionofcomplaintsandcourtorders. 8. All employees are expected to conduct their private business affairs in a manner that will not bring discredit to the company. The company will not serve as agent for the collectionofindebtednessclaimsagainstemployees.Therefore,onlythoseclaimsthat haveundergonedueprocessoflawwillbehonored.

- 48 -

EZLabManual COMPLAINTS 1. Complaintsagainstanemployeeforreasonsunassociatedwithhis/heremploymentby thecompanywillnotbeconsidered. 2. Telephonecomplaintsofcollectorsarenottobeknowinglyreferredtoanemployee. 3. Complaints,claimsorrequestsforcollectionassistancereceivedbythecompanyhead ormanagerwillbereferredtotheFinanceCommitteeforreply. POLITICALACTIVITY 1. Employeesmayparticipateinpoliticalactivityoutsidetheiremployment,butshallnot allow their interest in a particular party candidate, or political issue to affect the performance of their company duties, or interfere with other employees timely completionoftheircompanyresponsibilities. 2. An employee shall not, directly or by implication, use the name of the company or his/her affiliation with the company in the endorsement of political candidates, initiatives,referenda,orinsupportingoropposingapositiononotherpoliticalissues. 3. Anemployeewhoplanstocampaignforpublicofficeorwhoiselectedtopublicoffice should request a personal leave of absence, if his/her candidacy or public office prevents him/her from being available to perform his/her assigned duties and responsibilities. 4. The use of company resources, including mail, email or web sites, for the purpose of attempting to promote or defeat the election of a candidate for public office, or the passageoraballotmeasureisprohibited. SOLICITATION Solicitationsaretobediscouragedonthecompany,unlessanemployeeisactingfora charitable or service organization, and then such solicitation is not to be done during work hours of either the solicitor or the solicitee and with the understanding that the company reservestherighttocancelallsolicitationprivileges. DRESSCODE 1. Department heads shall have the right to prescribe certain reasonable standards of dress and appearance of their employees. These standards may be established to ensurethesafetyofemployees,tomaintainanappearanceofneatnessandcleanliness among employees, or to further other purposes related to conducting company business. 2. Departmentheadsthatwishtoestablishadepartmentaldresscodeareencouragedto consultwithaGeneralManager.Dresscodesmustbecommunicatedtoallemployees

- 49 -

EZLabManual LABORATORYSAFETY Inadditiontothehazardcommunicationtraining,alllaboratoryemployeesmustreceive annuallaboratorysafetytraining.Basiclaboratorysafetyrulesareasfollows. 1.Norunning,jumping,orhorseplayispermittedinlaboratories. 2.Allbiohazardous,hazardous,andradioactivematerialsmustbeproperlylabeledandstored. Useflammableandacidstoragecabinetsandexplosionproofrefrigeratorswhenrequired. 3.Stairways,hallways,exits,andaccesstoemergencyequipmentsuchasfireextinguishers, safetyshowers,andeyewashfountainsmustbekeptclear. 4.Noeating,drinking,applyingcosmetics,orsmokingisallowedinareaswherehazardous materialsarestoredorused. 5.Alwaysthoroughlywashyourhandsbeforeeatingorsmoking,oncompletionofwork. 6.Donotstorefoodordrinksinrefrigerators,freezers,orcontainersdesignatedforchemical, biohazardous,orradioactivestorage. 7.Compressedgascylindersmustbesecuredatalltimes,includingduringtransportandwhen empty.Cylindercapsmustbeinplacewhenthecylinderisnotinuse. 8.Donotworkaloneinalaboratoryiftheproceduresbeingconductedaredeemedhazardous bythelaboratorysupervisor.Assurethatatleasttwopeoplearepresentatalltimesifa compoundinuseishighlytoxicorofunknowntoxicity. 9.Laboratorycoatsandotherprotectiveclothingworninthelaboratoryareaarenottobe wornoutsidethelaboratory. 10.Sandals,opentoed,oropenheeledshoesshouldnotbeworninthelaboratory. 11.Neverdisposeofahazardous,biohazardous,orradioactivesubstancedownthedrainorin thetrashunlessyouhavebeenspecificallyauthorizedtodoso. 12.Radioactivematerialusageareasandanimalfacilitiesmusthavecontrolledaccessthatis strictlyenforced.Laboratoryareasshouldnotbeleftunattendedunlesstheareahasbeen secured. 13.Reportanyaccidentalexposure(inhalation,ingestion,skincontact,orinjection),injury,or spillstoyoursupervisorimmediately. - 50 -

EZLabManual CLOTHING Unlesstheworkeriswearingalaboratorycoat,looseortornclothingshouldbeavoideddueto thepotentialforignitability,absorption,andentanglementinmachinery.Danglingjewelryand excessivelylonghairposethesametypeofsafetyhazard.Tightringsorjewelrywhichcannot be easily removed should be avoided because of the danger of corrosive or irritating liquids producingskinirritation. Laboratorycoatsshouldbewornatalltimesinthelaboratoryareawheninfectiousorchemical hazardsarepresent.Laboratorycoatsandotherprotectiveclothingworninthelaboratoryarea shouldnotbewornoutsidethelaboratory. Shoesmustbewornatalltimeswhenworkingwithhazardousorinfectiousmaterials.Sandals, opentoedshoes,orperforatedshoesshouldnotbeworn. Gloves Manyhazardousmaterialscanbeabsorbedthroughtheskin.Others,suchasorganicsolvents candissolvethenaturalprotectiveoilsontheskin,leadingtochappedandcrackedskinandthe possibility of infection. Therefore, protective gloves must be worn when a potential skin exposureexistsorwherethereisapotentialforaccidentalspillsorcontamination.Besureto selecttherightgloveforthetaskathand. There is no glove currently available that will protect against all chemicals. Read the label, MSDS, or literature provided by the glove manufacturer to ensure that the proper glove has beenchosen. Before each use, gloves should be inspected for discoloration, punctures, and tears. If the gloves become contaminated, they should be removed and discarded as soon as possible. Always wash your hands immediately after removing gloves. Always remove gloves before leaving the immediate work site to prevent contamination of door knobs, light switches, telephones,etc.

- 51 -

EZLabManual

Laboratory StandardOperatingProcedures

- 52 -

EZLabManual PROTOCOLS a. Specimenhandling I. Receivingtoincludewalkinandsendinspecimensfromotherlabs. 1. Thespecimenisreceivedbythelaboratorystafffromthe receptionist.Itshouldbealwaysaccompaniedbyarequest.The requestingdataareenteredinthelogbook.Itshouldincludethe completenameofpatientthefirstname,middlename,surname, age,birthday,sexandOfficialReceipt(OR)Number. 11. Recording Resultofexaminationsarerecordedintheircorrespondinglogbooksaswell asonlaboratoryresultforms. 111.Rejectioncriteriaforadequacyandinadequacy a. Specimenrejection. 1. AllquantitynotsufficientspecimensQNS 2. ForUrine1hourofcollectionaftervoiding,itshouldbe30minutesafter voiding. 3. Stoolafter1hourofcollectionisrejected 4. Hemolyzedserumforelectrolytesexaminationandchemistry examination. 5. CRITERIAOFSPECIMEN ADEQUACY INADEQUACY SPECIMEN Urine approximately5ml lessthan2ml Feces Blood marblesize freshdroporgoodsmear verynegligibleamount toothin,withridges Wavesandholessmear

IV.REPEATTESTING 1. Repeattestingisdoneupontherequestofthephysician. 2. Whenerrorsignalsaredisplayedintheapparatusthetestis repeated. 3. Whenresultsareabnormallyhighorlowrepeattestingisdone.

- 53 -

EZLabManual IV.RELEASINGOFRESULTS 1. DoublecheckingofresultisdonebytheRMTbeforereleasingtothe client. V.SENDOUT 1. Examination that are not available in our laboratory are send to MT. GILEADpickupbytheirmessenger. 2. Histopath and papsmear are picked up by the secretary of pathologist daily. 3. Allsendoutresultsarealsoenteredinthererespectivelogbooks. VI.COURTESYREQUEST 1. If ever there is any courtesy request it will be known and address to the ownerforapproval. 2. Allcourtesyorpakishouldbeapprovedbythemanagementbeforeitwill bedone. VII.REFERRALSTOHEADOFLABORATORY. 1. Anybreakdown,malfunctioningandrepairsofmajorapparatusand equipmentsarealwaysreferredtothehead. 2. Whenthereisemergencyrunoutreagentorsuppliesrequestismade fromotherlaboratories. b.Procurement I.Supplies 1. Anorderedslipofsuppliesisaccomplishedbythepurchasing, fortheapprovaloftheheadthenpriortobookingbythesale representative. II. Reagents 1. An ordered slip of supplies is accomplished by the purchasing for the approvaloftheheadpriortobookingbythesalerepresentative. 2. Inventoryofallstocksofreagentsisupdatedregularly.

- 54 -

EZLabManual WASTEMANAGEMENTPROGRAM

- 55 -

EZLabManual

- 56 -

EZLabManual D.HANDLINGOFCOMPLAINTS,LABORATORYACCIDENTSANDINCIDENTREPORTS. 1. Allcomplaintsmaybeaddressverballyorwithwrittencommunicationtothe head. 2. Aninvestigationwillbeconductedtoconfirmthevalidityofsuchcomplaint. CONTINGENCYPLAN EQUIPMENTBREAKDOWN 1 Incaseofmachinebreakdown,MedicalTechnologist/swhousesthemachine dailymustdothepreventivemaintenancelikecleaningandcalibration. 2 If the Medical Technologist/s failed to sort out the problem of the machine, he/shemustcallanauthorizedtechnicianorserviceengineer. 3 Backupmachineshouldbeusedwhilethemachineisunderrepair. 4 Forthetestsandexaminationswithoutbackupmachines,testsmustbesentout assoonaspossibletolaboratorythatprovideshigherservicecapabilities.

- 57 -

EZLabManual

DIAGNOSTICTESTPROCEDURES

- 58 -

DIAGNOSTICTESTPROCEDURES

EZLabManual

MICROSCOPY URINALYSIS A. PhysicalExamination Takesnoteofthecolorandtransparency(usingcombitixteststrips). B. ChemicalExamination 1. Usefreshurinethathasnotbeencentrifuged.Thoroughlymixtheurinesample. 2. Briefly(nolongerthan1second)diptesturineonthebrimofthevessel. 3. After 60 seconds compare reaction color with the color scales on the label. Colorationsappearingonlyalongtheedgesofthetestpatches,ordevelopingafter more than 2 minutes, do not have any diagnostic significance, the urine specimen should not stand for more than hours before testing. Report: ph, protein, glucose, specificgravity The only chemical examination included in the routine analysis are for the determination of albumin and sugar. If we suspect by the color of the urine the presenceofbilepigmentsorifthecliniciansorequests,weexamineforthepresenceof bilebytheuseoftheSmithsmethod.TourineinaKolmertube,weadd35dropsof Gramsiodineandabluishgreencolorringatthelineofcontactindicatesthepresence ofbiletheoriginofwhichisthetotaldestructionofRBC.Theprincipleinvolvedinthis testisoxidationofthebileacidandwiththeformationofaseriesofcolorderivatives likebilirubin,biliverdin,etc.Wealsodothefoamtest.Positiveforbilearebigbubbles whichareyellowincolor. A. Albumin Albuminispresentinminutetracesintheurine(75mgm/houroutput)sosmall in amount that it could not be detected by any laboratory method or procedures. Differentlaboratoriesusesdifferentmethodsdetectingthepresenceofalbumininthe urine, but whatever test is performed, there is only one principle involved and that is coagulationbyheatandprecipitationbychemicalagents. Inourlaboratory,weuse10%aceticacidinHeatandaceticacidtest.Toacentrifugedurine, theupperthirdofwhichisheatedoveranopenflametoagentleboiling,thenadd35drops of10%HAc,andusingadarkbackground,allowingthelighttoenterwithintheurineandthe background.Noteforanycloudinessthatisformed.Anycloudinessthatformeduponheating and which disappears upon the addition of the acid is due to phosphates only. If it is really albumin, the cloudiness that formed upon heating will not disappear but will even be more marked. - 59 -

EZLabManual Therearemanyclassesofalbuminuria(presenceofexcessalbuminintheurine butallofthemfallsunderoneofthesethree(3)classes: 1. Physiologic Albuminuria where albumin is heavy, but which under the microscope,therearenosignificantsedimentsandwhichmaynotbedueto food rich in proteins taken or due to posture or exercise etc., and which is notpermanent. 2. AccidentalAlbuminuriawherealbuminisheavybecauseofthepresenceof albumnioussedimentslikepus,RBC,etc. 3. Pathological Albuminuria where the clinician are most interested in and weareapttoseeplentyofcasts.Thetwomostimportantproteinfoundin urineinpathologicalconditionsareserumalbuminandserumglobulin. B. Sugar There are various sugars found in the urine like lactose (usually found duringlactation),fructoseorlevilose(fruitsugar),galactose,dextroseorglucose (kind of sugar found in the urine of diabetics). These sugar are so small in amount that they cannot be detected by laboratory procedures. There are several methods for the determination of sugars in the urine but one that we adoptisthatonewhichismostcommoninlaboratoriesinthePhilippinesisthe Benedictstest,aqualitativetestforsugar. Thetestispopularduetoitssimplicityofapplication,sensitivenessand freedomoferror.Thisdoesnotreactwithnormalsugaroftheurinebutdetects whatever is excess of this amount as low as 0.2%. The principles involves in mostly all test is reduction. When the whitish blue cupric hydroxide in suspensionisheated,itisconvertedthesolublecuprichydroxidebutifsugaris present,thisisreducedtoinsolubleyelloworredcuprousoxidedependingupon theconcentrationofthesugarpresent. ThechemicalcompositionoftheBenedictsqualitativereagent: 17.3gms. Coppersulfatedissolvedinaround60cc.ofwater 173gms. SodiumCitrate 100gms. Sodiumcarbonate,ifahydrous 117118gmsifmonohydrated 200gramsifcrystalline These two chemicals are dissolved together with the aid of heating in around 500 cc. of distilled water. When all chemicals are dissolved, add the copper sulfate to the sodium citrate and carbonate and fill with distilled water up to 1,000ml.

- 60 -

EZLabManual PROCEDURE: Toa5cc.ofBenedictsqualitativereagentsplacedinaKolmerstube,add.5cc ofurine,mixedandplacedinboilingwaterfor5minutes.Letcoolandobserve forthecolorchange.Nochangeofcolorisnegative.Agreencoloris+;a++hasa yellowcolor;a+++assumesanorangecolorandabrickredisreportedas++++.

In our laboratory, we reduce the amount of the reagent to 1 cc. with a corresponding decrease in the quantity (0.1 cc.) of urine to be tested. This change in the amountdoes not make any difference in the endresult because evenifwereducetheamountofthereagent,wealsoreducethequantityofthe urineadded. C. MicroscopicExamination: Castsidentitytype.Reportnumberonslideoraverage.Lpf. WBC&RBCAveragerange/hpf. Epithelialcells,crystals,amorphousurates,mucusthreadsReport/lpf. Bacteriareport/hpfinpluses.

MICROSCOPY

PREGNANCYTEST

Pregnancy is a term used for a laboratory examination for the confirmation of early normalpregnancy,evaluationofabnormalpregnancy,andthestudyoftrophoblasticdisease. Allpregnancytestsaredesignedtomeasureordetecthumanchorionicgonadotropin{HCG},a hormoneproducebyviableplacentaltissue. IMPORTANCEOFPREGNANCYTEST: 1. Todetermineifawomanispregnantornot,becausesomewomanneedsmedications. 2. Todetermineiftherehasbeencompleteorincompleteabortion. 3. To differentiate pregnancy from uncommon condition like hydatidiform mole and chorioepithelioma. 4. Todifferentiateotherpelvicdiseases. 5. To differentiate pathologic condition of mole, like chorioepithelioma, teretoma, choriocarcinomaoftheovary.

- 61 -

EZLabManual PROCEDURE: 1. Instructpatienttocollect5mlofurineintoacleanvialsorcontainer. 2. Makesurethatthespecimensubmittedshouldbeclean;otherwise itmustbefilteredorcentrifuged. 3. Openthekitandusingtheencloseddropper,put3dropsofurineat thecirclering. 4. Waitfor3minutes,thenreadresults. INTERPRETATIONOFRESULTS: POSITIVEtwodistinctlineappearsonboththeTandCwindow. NEGATIVEonlyonelineappearsontheCLINE. MICROSCOPY SEMINALANALYSIS CollectionofSample Patientsshouldbeaskedtoabstainforthree(3)daysbeforesamplecollection.The mostsatisfactorysample,isthatcolletedintheclinicallaboratory.Ifthisisimpossible,patients itfromextremesoftemperature.Itshouldbecollectedinwidemouthplasticorglass containerbutnotincondomswhichmaycontainspermatozites Analysis 1. LIQUEFACTION.Freshlyejaculatedsemenisaviscid,opaque,whitecoagulum. Incubatethesampleat37Cfor30mins.Andexamineforliquefactionofthesemen. Normalsemenshouldbeliquefiedcompletely. 2. VOLUME.Normallythevolumeofseminalfluidrangesfrom1.5to5mlwithameanof 3.5ml.Infertilemalesoftenhaveanincreasedvolumeofseminalfluidwithadiminished spermcount. 3. pH.NormallythepHdoesnotvarymuchanditisusuallybetween7.7and8.0Values below7.0areusuallyassociatedwithsemenconsistingprimarilyofprostaticsecretion. 4. SPERMMOTILITY.Toevaluatemotility,placeadropofsemenonaslidethathasbeen prewarmedto37C.Examineunderx40objectiveanddeterminethepercentageof motilesperm.Normallymorethan70%ofspermatozoashowactivemotilitywithinour hourofejaculation. 5. SPERMCOUNT. a. Pipette0.02mlofwellmixedseminalfluidinto0.38mlofdilutingfluidinatest tube. b. Mixandchargeacountingchamber.Allowtosettlefortwominutes.

- 62 -

EZLabManual c. Counttwoofthelargesquares(2sqmm)andmultiplythefigureobtainedby 100,000toobtainthenumberperml.Normalvaluesrangesfrom60to150 millionperml.ofseminalfluid.Valueoflessthans20millionperml.are associatedwithinfertility. TheDilutingFluidin1%Fomalin 1. PhysicalExamination ColorOpaquefrayorpearlytolightyellow. Odoradistinctodor.Mustly,spermatic,fishy,chlovoxlike Viscocityfreshlyejaculatedspecimenishighlyviscid,ofcreamygelatinous, absenceofthischaracteristicsisanindicationoflowcount. MICROSCOPY MALARIALSMEAR Inidentificationofspeciesweusetwokindsofsmear: 1. Thicksmearusedinsearchingforbloodparasites 2. Thinsmearusedforstudyingmorphologyofparasites. Itisbesttoexaminebloodwhichistake1012hoursafterthechillto severalhoursbeforethechill. PreparationofStainingoftheThickSmear: 1. Touchthesurfaceoftheslideofthecrestofthelargedropofthebloodwithout touchingtheskin.Movetheslideinanarrowcircleintheblooduntilthesmear aboutthesizeofadimeismade. 2. Laytheslideflattodryatroomtemperatureandprotectagainstbrightsunlight, dust,andinsects. 3. Donotfix.Placetheslideonthestainingbridgeandgentlycoverwithdiluted Giemsasstainfor3minutesoruntilthehemoglobinisdissolvedfromthe erythrocytesmakingthemtransparentforexamination. 4. Thecytoplasmtothemalarialorganismstainblueandthechromatinispurplishred. Thengentlytilttheslideandallowtostaintodrainoff.Coveragainwithdiluted Giemsasstaintakingcarenottodislodgethedropofblood.Stainfor30minutes. 5. Removethestainbygentlytitlingtheslide.Washthestainwithneutralwatersoas nottoremovetheblooddropsfromtheslideandallowtodry.Examineunderoil immersionobjective. IdentificationofSpecies: 1. Itisbesttoexaminethebloodtakenduringtheperiod1012hoursafterthechillto severalhoursbeforethechills. - 63 -

EZLabManual 2. RegularsmearsaremadeandstainedlongerthantheregularstainingwithGiemsa. Thinsmeararebetterforstudyingthemorphologyoftheparasites. Bloodplatelestlyingontheerythrocytesmustnotbemistakenformalarial parasites. 3. Thicksmears a,Arelativelylargerquantityofbloodisplacedinasmallerareainaslideand stainedwithGiemsassothatthehemoglobinisdissolvedfromtheerythrocytes makingitsufficientlytransparentforexaminationwithtransmittedlight. b.Theorganismstainsthesameasinthinfilmsbutdonotappearin erythrocytes;onlyparasites,leukocytesandplateletsarestained. Procedure: 1. Fixbloodsmearwith95%absolutealcoholfor1minute. 2. Decantalcohol.Dontwashwithwater. 3. StainGiemsaworkingstandardsolutionfor7minutes. 4. Washwithwater,dryandexamine. Workingsolution(1:10dil.) - 1cc.stockGiemsa - 9cc.Buffersdistilledwater FECALYSIS MICROSCOPY A. MacroscopicorPhysicalExaminationTakenoteof: Form,consistency,color,odor,mucous,blood,parasites B. MicroscopicExaminationReportthefollowingbycountperlowpowerfieldor highpowerfield. Epithelialcells mucusthreads crystals Leukocytes tissuefragments Erythrocytes Bacteria Parasiteovareportnumber/coverslip FECALYSIS:NORMAL FormConsistencySoftorsemiformed ColorLightbrowntodarkbrown OdorOffensivebutnotexcessivelyfoul Mucousverysmallamount Bloodnegative Parasitesnegative Buscellsveryinsignificantamount - 64 -

EZLabManual Undigestedfoodspresent(butthosethatarereallyandnormallycannotbe digested)

Fecesarenormallycomposedofwaterandsolids,suchasbacteria,intestinalconcretions, foodresiduessuchasseedsvegetablesskins,hairs,fibers,vegetablecellsmusclefibers,fat dropletsandothersolublesubstances. Importanceoffecalexamination: a. Presenceofaspecificdiseaseagentsuchasintestinalparasites. b. Indirectevidenceofmalfunctionofsomeportionofthegastrointestinal trac,theliverorpancreas. CollectionofSpecimen: Patienttoundergofecalexaminationshouldavoidtakingbarium,bismuthoreilfor5days precedingtheexamination. 1. Passoutinbedpanoranysterledcontainerandavoidadmixtureofurine,dirt,cotton fibers,tissuepapers,gauzethreads. 2. Bymeansofamatchstickortoothpickorapplicatorstickortonguedepressorget aboutapeasizeormarblesizeorthumbsizeorguavasize. 3. Themidportionisthebest,however,selectalsothebloodyormucoidportion. 4. Incaseofliquidstools,takeabout10cc.of1/3fullofaneighttotenouncebottle container. 5. Containeritispreferabletohaveaclean,dryandsteriledcontainerofawidemouth withascrewcaporaparaffinizedboxoficecreamofaboxofabout28ouncesinsize. 6. Examineimmediately,ifdelayednotethefreshspecimenscolor,odorandconsistency. Preservatives: a. refrigerator b. 2%5%,10%formaline c. 40%alcohol d. Freezingindryice e. Bufferedglycerolsalinesolution f. MIFpreservative(MerthiolateIodine) g. PVAfixative(Polyvinylalcohol) Proceduresinpreservationofstoolspecimen: a. Ifthespecimenisformedorsoftandenoughwaterandmixitthoroughlywith applicatorstick. b. Strainoffilteritthrougha2layeredgauzeofcheesecloth c. Leavethestrainedspecimenfor30mins.To1hourtoallowsettlingof sediments. - 65 -

EZLabManual d. Decantthesupernatantliquidandreplaceiteiththepreservativeyouwouldlike toadd. e. Ifstooliswateryorliquid,specimenissimplystrainedthrough2layeredgauze orcheeseclothanddotheotherproceduresasabove.

MacroscopicExaminationsofPhysicalExamination: 1. Amountandfrequency Normally100to200gramsperdayand1to2timesaday250gramsperdayincaseof moistandvegetablediet1/3toofthedryweightisbacteria. 2. Form,Consistencyandsize Normallysoftorsemiformed Abnormala.soft,mushyandwaterystoolsarefoundindiarrheaoroftertheuse ofcathartics. b.Excessivelyhardandscybalousstoolsarefoundinconstipation c.Flattenedorribbonlikestoolsarefoundinspasticcolitis,stricture, tumor,ulcerorcarcinomaandobstructionofthelowerportionofthecolon. 3. Blood Normallynobloodisfoundinthestool Abnormal A.GrossBlood Takenotethecharacterofthestool,iftarry,reddishorbrightred,alsonote thedistributionofthebloodwhetherevenlymixed,partiallymixedoronly ontheoutside. 1. Ifstoolisdarkincolorortarryandbloodisthoroughlymixedtheoriginof thebloodisusuallyfromtheuppergastrointestinaltractandeveninthe respiratoryorganssuchasthenoseandmouth. 2. Ifthestoolconsistofbrightredbloodandnotthoroughlymixedthe originofbloodismostlikelyfromthelowedbowelandanalopening. B.OccultBlood Thisisundetectedbythenakedeyenadcanonlybedetectedby chemical. 4.Parasites Normallythereshouldbenoparasitesinstool. Abnormallysomeparasitescanbeeasilyseenliketheadultascarisand segmentsoftapeworms.Otheradultsparasitessuchashookworms, - 66 -

EZLabManual Pinwormsandtrichinellacanonlybeseenbysedimentationprocedures Andmixingwithwaterandrunningthroughafinesieveorgauze.Ahandlens maybenecessarytoidentifythetiniestones.Ovum,cystandtrophozoitesare beenbytheuseofamicroscope. ArtifactsMostLikelyMistakenforParasites: a. Poorlymasticatesceleryorgreensforroundworms. b. Cellsfromorangesforpinworms c. Fibersfrombananafortapewormsandova. 5.PusCells Normallyaveryinsignificantnumberofpuscellsispresent. Abnormallypresenceisindicateofulcerativecolitis,whereasmucus Aloneindicatesmucuscolitis.Generallypresentinulcerativecasesofthe bowelsduetosyphilis,tuberculosis,bacillarydysenteryorinlargequantities aftertheruptureofanabcess. 6.UndigestedFoods Normallythesearepresentbutthosethatarereallyandnormallycan Notbedigested.Thesearehowever,oflessimportance.Mostcommonly seenateseedsandfiverswhichareconfusedforconcretionsandparasites. Alsocurd,likemassesareoftenmistakenforpusandconcretions.Fatsmay beidentifiedbytheirsolubilityinether. 7. Concretions Normallyisnotseeninstool MICROSCOPY GRAMSTAINING PROCEDURE: 1. Heatfixedsmearstobestained. 2. StainsmearwithCRYSTALVIOLETfor1minute. 3. Washwithtapwater. 4. MordantwithGRAMSIODINEfor1minute. 5. Washwithtapwater. 6. DecolorizewithACIDALCOHOLuntilnomorepinkcomesoff. 7. Washwithtapwater. 8. CounterstainwithSAFRANINfor1minute. - 67 -

EZLabManual 9. Washwithtapwater. 10. Blotslidedry.Examineunderoilimmersion. Note:Filterstaindaily RESULTS: Gram(+)organismPURPLE Gram()organismPINK MICROSCOPY OccultBloodDetermination(BenzidineFilterPaperMethod(Modified) REAGENTS: FilterPaper ReagentA ReagentB Note:Keeptightlycappedandcoverimmediatelyafteruse. PROCEDURE: 1.Smearasmallparticleoffecesonapieceoffilterpaperwithawoodenapplicator stick. 2. Add3dropsofReagentA. 3. Add3dropsofReagentB. 4. Readwithin1minute. RESULTS: POSITIVEAnytraceofBLUEonorattheedgeofthesmearindicatesapositive testforoccultblood. NEGATIVENodetectableblueonorattheedgeofthesmearindicatesanegative testforoccultblood.

- 68 -

EZLabManual MICROSCOPY

CEREBROSPINALFLUIDEXAMINATION

A.MacroscopicorPhysicalExamination observationthefollowing:color,turbidity,coagulation,reaction,specificgravity B.ChemicalExamination PandysTestforAlbumin Reagents: Phenolcrystals 10gms. Distilledwater 100gms Procedure: 1. Place1ccofasaturatedaqueoussolutionofphenolinsmalltesttube. 2. Add1largedropofspinalfluid. 3. Abluishwhitecloudformingimmediatelyaroundthedropindicatedsan abnormalamountofalbumin. 4. Normalfluidsmayshowafainttracebutthisshouldbereportedasnegative. CEREBROSPINALFLUID MAGROSCOPICORPHYSICALEXAMINATIONS 1.AMOUNT a.Thenormalamountisroughlyestimatedtobeabout1cc.perpoundorbodyweight. Alsoroughlyestimatedtobeabout100to150cc.everydayproduction. b. there is increased amount in acute and chronic congestion of the meninges due to increasedtransmudationofplasmathroughthecapabilityofthechororoidplexus. c. There is also an increased amount in acute and dochronic infection due to the productionofinflammatoryexudaticandtoincreasedpermeabilityofthecapillaries. 2.PRESSURE a.Normalpressureforthehorizontalpositionvariesbetween70and200mm.ofwater (0.8mm.ofmercury)withtheaverageof100to150mm. 3.Color a.Normallythefluidisasclearandcolorlessasdistilledwater. b. Bright red due to fresh blood from a vessel punctured while inserting the needle; uponcentrifugationthesupermutantisclear. c.Dullredofbrown(dependingontheageofthelesion).Inhemorrhagsaguechronic hemorrhagicpachmeningitis.After48hoursmemolysisbegins,sothatahemorrhageof

- 69 -

EZLabManual atleastthisageisrecognizedbytheyelloworredcolorofthesupernatantfluidsafter contrifugation. d. Yellow (xanthochromic) may be to blood pigments resulting from disintegration of RBCwithinthesubrachnoidspaceortoalteredpermeabilityoftheliningmembranesto pigmentsinthebloodplasmawhichundernormalconditionareexcluded. e.Greenishorgrayishduetopuscellsinsevereinflammatoryreactions. 4.TURBIDITY a. In acute meningitis the fluid may exhibit varying degrees of cloudiness, from slight turbiditytothecapacityofalmostpurepus. b.Inthelessacutestageofepidemicmeningitis,itissometimesquiteclear. c.Itisusuallyclearintuberculosisandsyphiliticmeningitis,tabes,andpolimyclitis. 5.COAGULATION a.Normalspinalfluiddoesnotcoagulated b.Abnormal 1.Thefluidclotswhenthereisanincreaseinproteinincludingfibrinogen 2.Numeroussmallcoagulaoccurinpasesis 3.Acobweborpinetreedelicatedcoagulumistypicaloftuberculosismeningitis;it formsonthesurfacefluidsandextendsdownthemiddleofthetube. a)Twelveormorehoursmayberequiredforitsformation b)Theabsenceofpellicledoesnotexcludetuberculousmeningitis. 4.Heavycoagulumandsedimentoccurinacutecuupurativemeningitis. 5. Complete coagulation with xanthochromia without hemorrhage occurs in Froins syndrome(spinalsubarachenid)block 6.Reaction Thenormalphis7.4to7.5 7.SpecificGravity Thenormalspecificgravityis1.003to1.008 CHEMICALEXAMINATION 1.ProteinsQualitativeTest GeneralConsiderations a. Theproteinofchiefintrestisglobulin b. Thetestforglobulinisvaluelesswhenappliedtofluidcontainingblood,owingtothe presenceofseruminglobulins. c. Ifthefluidiscloudy,itshouldbecentrifugedandtheclearsupernatantfluidusedfor thetest. A. NoneApeltTestforGlobulin Reagents: Merckspurifiedneutralammoniumsulfat.85gms. Distilledwater100cc. - 70 -

EZLabManual Boilandfilteredassoonasitiscooled. PROCEDURE: a. Placeequalamountofspinalfluidandsaturatedsolutionofammoniumsulfateina testtubeandmixbyinvertingthetube. b. Allowthetubetostandforthreeminutes. c. Fluudthatcontainsanormalamountofglobulinwillremainclearorbecomeslightly opalescent,ifanexcessofglobulinispresentacloudyprecipitatewillform. B. RossJonesTest a. PlaceLcc.ofspinalfluidintoasmalltesttube b. Add1cc.ofammonounsulfatetothebottomofthetube(plugthepipetand immergetothebottomofthetubeandreleasetheammoniumsulfate,the plugitagainandwithdrawthepipet) c. Aclearout,thin,grayishwhiteringappearsatthezoneofcontractfor positivereaction.Undernormalconditiontheringwillappearafter5 minutes. d. Observefor3minutes,thenmixthesolution e. Interpretation: - Ringin3minutesandnotraceaftermixing. - Ringin3minutesandfaintopalescentaftermixing. - Ringin3minutesanddefinitecloudaftermixing. - Ringin3minutesandheavycloudaftermixing. ForAlbuminTest a. Shakethecontentofthetubeusedintheglobulintestandfilter b. Aciditywith1dropof10%aceticandboil c. Alightcloudinessisnormal d. Interpretations: Negativeaslightcloudiness - definitecloudinesswithfireprecipitate - aflocculentprecipitateinaslightlycloudyfluid - aheavyflocculentprecipitateinaclearfluid. C. PandysTestforAlbumin Reagent Phenolcrystals10gms. Distilledwater100gms. Procedure: a. Place1cc.ofasaturatedaquecoussolutionofphenolinasmalltesttube. b. Add1largedropofspinalfluid c. Abluishwhitecloudformingimmediatelyaroundthedropindicatesan abnormalamountofalbumin. d. Normalfluidsmayshowafainttracbutthisshouldbereportedasnegative. - 71 -

EZLabManual TotalProteinsQuantitative(Normalvalueis30mg.per100cc.) Principle: The protein is precipitated from the spinal fluid with acid tungstate, dissolved, and reprecipetated.Thenitrogenisdeterminedonanaliquietportionafterbeingputintosolution withtheaidofsodiumhydroxide. GeneralConsiderations
a. Presencesofbloodgivefalsehighvalues. b. Presencesofbacteriagivesunreliableresults c. Delayofanalysisincreasesthevalueobtainedunlessfluidiskeptsterileandtightly corked.

ProcedureofthetestissimilartoBloodChemistryprocedure. HEMATOLOGY ACIDFASTSTAINING PROCEDURE: Heatfixedsmearstobestained. FloodsmearwithCARBOLFUCHSINandsteamfor5minutes. Washgentlywithtapwater. DecolorizewithACIDALCOHOLonlyuntilfaintpinkcolorremains. Washwithtapwater. CounterstainwithMETHYLENEBLUEfor1minute. Washgentlywithtapwater. Blottheslidedry. Examineunderoilimmersion. RESULTS: ACIDFASTBACILLIREDORDARKPINK NONACIDFASTORGANISMBLUE

- 72 -

EZLabManual HEMATOLOGY PLATELETCOUNT IndirectMethod 1. Make a good blood smear from venous blood taken into EDTA, or from a freely flowingskinpuncture.Dryintheairandstainwithrightshemaquickstain. 2. Usinganoilimmersionobjective,countthenumberofplateletsper1000RBCs. 3. Performanaccurateredcellcount. 4. Calculate as follows: If P is the number of platelets counted per 1000 RBCs, then platelets =PxRBCsperul 1000 Alternatively,theformulacanbesimplifiedasfollows: Plateletscountx109/L=PxRBC Normalvalues:150,000350,000/mm ReesEcker 1. FillaredcelldilutingpipettewithfreshlyfilteredReesEckersolutionandexpelit, leavingafilmofthesolutioninsidethepipette. 2. Drawcapillarybloodtothe0.5markontherbcpipette. 3. Wipeexcessbloodanddiluteto101markwithReesEckerdilutingfluid. 4. Shakepipetteforseveralminutes.Dischargeafewdrops,thenchargeboth chambersofthecountingchamber. 5. Allowthethrombocytestosettlefor15minutes.Topreventdrying,placethe countingchamberonadampfilterpaperinsideaPetridish. 6. Countthefourbigsquaresasinwbc. 7. Calculateasfollows: No.ofcellscountedx2000=platelet/cu.mm NormalValue=150.000350,000/cu.mm HEMATOLOGY COMPLETEBLOODCOUNT A. WHITEBLOODCELLCOUNT Equipmentandreagents: 1. Whitebloodcelldilutingfipette 2. Hemacytometer 3. Hemometerwiththickcovership 4. Microscope 5. Cottonballswithalcohol - 73 -

EZLabManual Dilutingfluid: GlacialAceticAcid 1cc. 1%Crystalviolentsolution 1cc. Distilledwater 100cc. RulesincountingLeukocytesorWhiteBloodsCells 1. Discard23dropsofdilutedblood(suckedatexactly)0.5markofthepipetteand diluted up to 101 mark past the bulb) from the pipette to the hemacytometer, beforechargingshakethepipettefor23minutes,placethecoverglassIitsproper positionovertheruledareaofthehemacytometer. 2. Thepipetteisheldpracticallyhorizontalwhilefillingthecountingchambertokeep tomuchfluidfromrunningunderthecoverglass. 3. Allowoneminutesfortheleukocytestosettle,focusunderlowpower. 4. Ifthereisdoubtastowhethertheobjectisleukocyteshavedefinitecelloutlineand welldefinednuclearstructure.Plateletappearsasverysmallrefractilebodieswith nuclei.Erythrocytesshouldnotbepresentbecausethedilutingfluidhemolyzethem. 5. Count4cornergroupof16squares..countfirsttheupperleftsquare,thenupper rightdropdowntothelowerrightandlasttothelowerleft. 6. Calculation: Cellscountedx20(dilutionx10(depth) No.ofsquaremm.counted(4sq.mm) =leukocyte/cu.mm. Orcellscountedx50=leukocyte/cu/mm B. HEMATOCRITDETERMINATION 1. Fillcapillarytubingwithblood 2. Sealcapillarytubing. 3. Centrifugecapillarytubingwithbloodfor34minutes 4. Read the volume of packed red blood cells cover the graduated plastic scale. Readatthemeniscusoftheerythrocyteoftheerythrocytecolumn. 5. Thereadingrepresentsthevolumeofpackederythrocytesper100cc.ofblood. C.Hemoglobin Method:cyanmethemoglobin Among the forms of hemoglobin well adapted to photometry, Cyanmethemoglobinhasoutstandingadvantages. a. Allformsofhemoglobinlikelytobeformedinbloodwithexceptionof Sulfomethemoglobin, are quantitatively converted to cyanmethemoglobinupontheadditionofasinglereagent. b. Solution of cyanmethemoglobin are the most stable of the various hemoglobinpigment.

- 74 -

EZLabManual 1. Drabkins Procedure: 1. Measure5mlofdrabkinssolutionintoatesttube. 2. with a Sahli pipette, measure 20 cumm blood and transfer into the test tube containingDabkinssolution. 3. Mix,letStandfor10minutestoallowformationofcyanmethemoglobin. 4. UsingStatFaxmachineFilterat550readtoobtainresult. D.DifferentialCount MakingtheBloodSmear(TwoSlideMethod) Procedure: 1. Prepareasmalldropofbloodinthemiddlethefirstatanofoneoftheslides. 2. Holdthesecondslide(spreader)abovethefirstatanangleof3045degrees. 3. Drawthespreadertowardthedropofbloodandcontractthebloodtheblood shouldfanofspreadouttotheedgesofthespreader.Ifitdoesnotfan,wigglethe spreaderalittletomakeitdoso.(Dontletthebloodgetaheadofthespreader becauseitmaycutordistortthecells)/ 4. Pushthespreadersmoothlyandrapidlyovertheentirelengthoftheslideat constantangle(3045)degreesandkeeptheedgeofthespreaderpressedfirmly againsttheslide. 5. Airdrybeforestaining. HEMATOLOGY STAINING(UsingHemaQuickStainSet) E.StainingReagents: HemaQuicksolution1250mL HemaQuicksolution2250mL HemaQuicksolution3250mL HemaQuicksolution4250mL HemaQuicksolution5250mL NOTE:HemaQuickStain1250mL(ForRapidStainingofbloodSmoars) Thesesolutionsshouldbeplacedincoplinjarsorotherdipstainingdevicesthat canbecoveredtopreventevaporation.

- 75 -

EZLabManual PROCEDURE: 1. Prepareslidesinthesamewayyouarepreparingslidesfordifferential cound. 2. Airdrysmearbeforestaining. 3. Dipairdriedsmearfive(5)timesforaboutone(1)secondeachdipinto Solution1,Solution2,andSolution3. 4. RinsetheslideintoSolution4. 5. Allowtodry. 6. Examineunderoilimmersionlens. RESULTS: ERYTHROCYTES pinkishcolor PLATELETS purplegranules POLYMORPHONUCLEARNEUTROPHILS NUCLEUS darkblue CYTOPLASMpalepink GRANULES reddishpurple HEMATOLOGY RBCCount: Men4,500,000/cu.mm. Women4,000,0005,000,000/cu.mm Normalwomeninlatepregnancy3,000,000 5,000,000/cu.mm. WBCCount:Male/Female5,00010,000/cu.mm. Heloglobin:Men1418gm/100cc Women1216gm/100cc Children1426gm/100cc Hematocrit:Men4054vol.%Ave.47vol.% Women3747vol.%Ave.47.% NormalWomeninlatepregnancy2324vol.% CoagulationTime:Slidemethod24minute PlateletCount: 250,000300,00/cu.mm.

- 76 -

EZLabManual HEMATOLOGY COAGULATIONTIME MethodSlideMethod Procedure: 1. Makeadeeppunctureintheearlobeortheringfinger 2. Thesizeofthewoundshouldbesuchthattheblotofdiameters.Notethetimeof thefirstdropbloodthatappears. 3. Removethedropsofbloodwiththefilterpaperat30secondsintervalsbeingcareful nottotouchtheskin.Theendpointisreachedwhennomorebloodappears. 4. Ifmoderatelyprolongedthe20thdropwillbeofthesizeofthefirstandif markedlyprolonged,the20thdropwillbeaslargeasthefirst HEMATOLOGY BLEEDINGTIME(DukesMethod) PROCEDURE: 1. Makeadeeppunctureintheearlobeortheringfinger. 2. Thesizeofthewoundshouldbesuchasinblotofthebloodafter30seconds intervalsis12mm.indiameter.Notethetimeofthefirstdropofbloodthat appears. 3. Removethedropsofthebloodwiththefilterpaperat30secondsintervalsbeing carefulnottotouchtheskin.Theendpointisreachedwhennomorebloodappears. 4. Ifmoderatelyprolongedthe20thdropwillbeofthesizeofthefirstandif markedlyprolonged,the20thdropwillbeaslargeasthefirst. HEMATOLOGY RETICULOCYTECOUNT RETICULOCYTES are proerythrocytes or pornormocytes or skin cell are which are immature erythrocytes which show reticulum when stained by some supravital method. This vital stain precipitatesthebasophilicsubstancesofthereticulocytes. USESOFRETICULOCYTECOUNT: 1. Asdiagnosisforleadpoisoning,hemolyticicterus. 2. TodetermineresponsesofperniciousanemiatoVit.B12 3. Todiagnoseacutehemorrhage. - 77 -

EZLabManual COMPOSITIONOFSOLUTIONAANDBFORRETICULOCYTECOUNT: A1%ofbrilliantcresylblue B1%Koxalate Reticulocytescountbyseweredsabinmethyleneblue. NORMALVALUES: 0.51.5% METHODSOFRETICULOCYTESCOUNT: 1. MethodofchoiceusingMethelynebluenormalsolution 2. Methodofcook,mayerandtureensmethod. 3. Rapidmethodofschilling. 4. StiffmodificationofKlempereemethod. 5. Sabinmethod. TECHNIQUEOFCOOK,MAYERANDTUREENMETHOD: 1. Placefivedropsof1%solutionofbrilliantcreyslblueinsalineinawatchglassoradd 1dropof1%solutionofPotassiumoxalateinsaline. 2. UsingSahlipipette,drawupsomebloodandequalamountofthemixedstin.To avoidprecipitate,donotuseexcessivestain. 3. Mixthoroughlybyalternatelyblowingoutintothepipette;allowstanding5or6 minutes.CoverwithaPetridishtopreventevaporation. 4. Placeadropofmixtureofbloodandstainupontheendsofaglassslideandspread inthemannerofmakingordinarybloodfilm. 5. Dryquicklytopreventdistortionofthecells.Thisslidemaybeexamined immediatelyorcounterstainedwitheitherWrightorGiemsasstain.IfGiemsais usedfixedfirstfor3minutesinMethylalcoholpreparatorytostaining. 6. Placeapieceofpaper,thesizeoftheeyepiecewithanopeningof5mm.squarecut. Thisreducethesizeofthefield. 7. Count1000erythrocytesnotingdownthenumberoffilaments,skeinsorwreaths. 8. Calculation= No.ofreticulatedcellsx100%reticulocytes orNo.ofreticulocytes 1000 9. Normalreticulocytescountis0.51.5% 10. Interpretation: Increase reticulocytosis is a sign of early regeneration or erythrocytes by the bone marrow. Count of less than 0.5% indicates an abnormally alow rate of productionorerythrocytesbythebonemarrow

- 78 -

DIAGNOSTICTESTPROCEDURES HEMATOLOGY PROTHROMBINTIMETEST(PTDETERMINATION)

EZLabManual

PRINCIPLE: TheonestageProthrombinTimemeasurestheclottingtimeoftestplasmaaftertheadditiono ThromboplastinreagentcontainingCalciumchloride.Thereagentsuppliesasourceoftissue trhromboplastin, activing Factor VII, and is therefore sensitive to all Stage II and III Factors. DeficienciesofStage1Factors(VII,IX,XIandXII)arenotdetectedbytest. PREPARATION A. Anticoagulant.Usebufferedsodiumcitrate3.2% B. PlasmaCollection 1. Obtainvenousbloodbycleanvenipuncture 2. Useonlycitrateplasmafortestanalysisruns. 3. Mix9partsvenousbloodwith1part3.2%(0.105M)sodiumcitrate. 4. Centrifugethemixtureof1500gforapprox.10minutes. 5. Removeplasmafromthetubewithin60min.usingaplasticpipette andstoreinaplastictube. 6. Test plasma sample within 4 hr, otherwise store frozen and thaw justpriortouse. C. ReagentReconstitution Reconstitutewithhighpuritywateratthevolumestatedontheviallabel.Letstandatroom temperature with occasional swirling until solution is complete. Stable for 7 days after reconstitutionwhenstoredat280C,avoidprolongedheating. D. CoatronM1Preparation 1. Turnoninstrumentandwaituntilthereadyledislightning. 2. Turnonprinterifconnected 3. SelectPTasactivetest 4. Checkcalibration 5. Allowreagenttoprewaratleast5minutes PROCEDUREONCOATRONM1 1. Pipette25ulplasmaintocuvette 2. Prewarmplasmafor1min 3. Transfercuvettetomeasuringposition 4. Activateoptic(presskeyOptic) 5. Add50u;prewarmedThromboplastinandsimultaneouslystarttheoptic,(press keyOpitcagain) 6. Theresultwillreadmaximal300secs.Ifnoclotisdetected,thedisplaywillread +++.+s 7. TheresultisdisplayedinsecondsandINR. - 79 -

EZLabManual ASSAYCALIBRATION ForINRcalibrationofPTtwoparametersarerequired. Thenormalvalue(determinePTnormalvaluewithCalibratornormal,expectedrangeois between11to14s) TheISIvalue(readISIvalueonthelabelofthereagentvial) EnterbothvaluesintotheCoatronM1. QUALITYCONTROL Controlplasmashouldbetestedinconjunctionwithpatientsamples.Itisrecommendedthatat leastoneNormalandoneAbnormalberunatleasteachshiftandaminimumofonceper20 patient samples. A Control Range should be established by the laboratory to determines the allowablevariationinadaytodayperformanceofeachControlPlasma. EXPECTEDRESULTSOFCONTROL
ControlNormal ControlA1 Control 2 A 0.81.3INR 2.03.5INR 5.08.0INR

BLOODCHEMISTRY PTTDETERMINATION The APTT test measures the clotting time of test plasma after the addition of APTT reagent, thenallowinganactivationtime,followedbytheadditionofcalciumchloride.Deficienciesof approximately40%andlowerofFactorsVIII,IXandXIIwillresultinaprolongedAPTT.Heparin, inthepresenceofadequateamountsofATIIIwillalsoresultinaprolongedAPTT. REAGENT APTTReagent CalciumChloride PREPARATION A. Anticoagulant.Usebufferedsodiumcitrate,3.2%. B. PLASMACOLLECTION 3. Obtainvenousbloodbycleanvenipuncture. 4. Useonlycitrateplasmafortestanalysisruns. 5. Mix9partsvenousvloodwith2part3.2%(0.105M)sodiumcitrate 6. Centrifugethemixtureat1500gforapprox.10minutes. 7. Removeplasmafromthetubewithin60min.usingaplastic npipetteandstoreinaplastictube. - 80 -

EZLabManual 8. Testplasmasamplewithin4hr,otherwisefrozenandthawjust priortouse. C. ReagentReconstitution Bring to room temperature prior to use. Mix well by swirling or inversion and maintainastirbarinreagentcontainerduringusetoavoidtheparticleactivator fromsettlingout.Avoidprolongedheating. CoatronM1Preparation 1. TurnoninstrumentandwaituntilthereadyLEDislightning. 2. Turnonprinterifconnected 3. SelectPTTasactivetest 4. AllowCaCl2toprewaratleast5minutes.

D.

ASSAYCALIBRATION PPTresultsshouldbenormalizedagainstanormalvalue,whichisobtainedfromaPTTof CalibratorNormal.expectedrange:seeboxinsertofCalibrator. EnterthenormalvalueintotheCoatronM1.ThePTTisthendisplayedinsecondsand normalizedintoRatio. PROCEDUREONCOATRONM1 1. Pipette25ulplasmaintocuvette. 2. Add25ulAPTTtoplasma 3. Incubateexactlyfor5minutes 4. Transfercuvettetomeasuringposition 5. Activateoptic(presskeyOptic 6. Add25ulprewarmedCalciumChlorideandsimultaneouslystartthetopic.(presskey Opticagain) 7. Theinstrumentwillreadmaximal300seconds.Ifnoclotisdetected,thedisplaywill read===.=s 8. TheresultisdisplayedinsecondsandRatio 9. QUALITYCONTROL Controlplasmashouldbetestedinconjunctionwithpatientsamples.Itisrecommendedthatat leastoneNormalandoneAbnormalberunatleasteachshiftandaminimumofonceper20 patientsamples.AControlRangeshouldbeestablishedbythelaboratorytodeterminethe allowablevariationindaytodayperformanceofeachControlPlasma. - 81 -

EZLabManual EXPECTEDRESULTSOFCONTROLS ControlNormal 0.81.2Ratio ControlA1 1.52.0Ratio ControlA2 2.03.0Ratio BLOODCHEMISTRY GLYCOHEMOGLOBIN SPECIMENCOLLECTIONANDPREPARATION: Special preparation of the patient is unnecessary. Fasting specimens are not required. Nospecialadditivesorpreservativesotherthantheanticoagulantsarerequired.Collectvenous bloodwithEDTAusingaseptictechnique.GlycohemoglobininwholebloodcollectedwithEDTA isstableforoneweekat28C. PROCEDURE: A.HEMOLYSATEPREPARATION: 1.Dispense500ulLysingReagentintotubeslabeled:Standard,Control,Sample 2. Place 100 ul of the wellmixed blood sample, standard or control into the appropriatelylabelledtube.Mixwell. 3.Allowtostandfor5minutes. Note:Plasticorglasstubesofappropriatesizeareacceptable. B.GLYCOHEMOGLOBINPREPARATION: 1.Dispense3.0mlofglycohemoglobincationexchangeresinintoglasstubelabeled Standard,Control,Sample. NOTE:Beforeusing,mixtheresinbyinvertingatleast10times.Swirlthebottleafter Additiontoeach5tubes. 2.Add100ulofthehemolysatefromstepA. 3.Positionthefilterseparatorsinthetubessothattherubbersleeveisapproximately 1cmabovetheliquidlevel. 4.Placethetubesontherockerorrotatorandmixcontinuouslyfor5minutes. 5.Removethetubesontherockerorrotator. 6.Pushthefilterseparatorintothetubesuntiltheresinisfirmlypacked. 7.Thesupernatantmaybepouredintoanothertubeordirectlyintoacuvettefor absorbancemeasurement. 8.Adjusttheinstrumenttozeroabsorbanceat415nmwithdeionizedwaterasthe blank.Wavelengthrange390420nm. - 82 -

EZLabManual EXPECTEDVALUES:6.68.6% ENZYMATICGLUCOSE(Trinder/GODPOD)

BLOODCHEMISTRY

INTENDEDUSE Forinvitrodiagnosticuseinthequantitativedeterminationofglucoseinserumorplasma REAGENTS: GLUCOSEBUFFER GLUCOSEENZYMEREAGENT GLUCOSESTANDARD(100MG/D1) SPECIMEN Plasma or serum should be separated from the cells as soon as possible after collection. Samplesshouldbeassayedpromptly.Glycolysisinserumsamplescancauseaglucoselossof about510mg/d1/hratroomtemperature(3). MANUALPROCEDURE
Allreagentsmustattainroomtemperature(2025oC)priortouse.Place2.5mlofGlucosebuffer into test vials labeled: Unknown, control, standard, blank Add 0.020ml (20ul) of sample to the corresponding vial. Add 0.100ml (100ul) of glucose enzyme reagent to all vials, mix by gentle swirling and return to the 37oC heating bath for exactly 10 minutes. (Alternative: 25 minutes at room temperature2025oC).Setthewavelengthofthephotometerat510nmandzerothephotometerwith theBLANK.Readtheremainingvials,recordtheabsorbanceandproceedtoCalculationwithExample. Note:Foradirectreadoutinstrument,setreadouttoconcentrationofstandard(100mg/d1).Readthe unknownconcentrationdirectly. ENDPOINTSTABILITY

Thefinalcoloredreactionproductshouldbemeasuredwithin60minutes.Avoiddirect sunlight. CALCULATIONWITHEXAMPLE WhereA=absorbance,U=UNKNOWN,S=STANDARD,C=concentration: PROCEDURELIMITATIONS: Expectedvalues(6) Adultfastingplasmaorserum: 70105mg/d1

- 83 -

EZLabManual PROCEDURENOTES Forglucoselevelsexceeding500mg/d1,reruntheprocedureona1:10dilutionofsamplewith 0.85%salineandmultiplytheresultby10. Formoderatelyictericorlipemicsample,add0.20ml(20ul)ofsampleto2.6mlof0.85% saline,mixandmeasureabsorbanceat510nmversus0.85%salineBLANK.Subtractthis absorbancevaluefromtheabsorbancevalueobtainedinitiallyintheaboveProcedure,and thencalculatecorrectedresults. QUALITYCONTROL Qualitycontrolserashouldbeusedroutinelytomonitortestprecision(11).Refertothe manufacturersproductinsertforanalytestabilityandacceptablelimits.SterlingNormal Control(Cat.#1003N)andSterlingAbnormalControl(Cat.#1003A)arerecommendedforthis purpose. BLOODCHEMISTRY ENZYMATICURICACID(ModifiedTrinder) INTENDEDUSE FORINVITRODIAGNOSTICUSEinthequantitativedeterminationofuricacidinserumor plasma. REAGENTS: Thefollowingreagentsshouldbestoredunderrefrigeration(28oC)andmaybeused untiltheexpirationdateindicatedonthebottles. URICACIDBUFFER URICACIDENZYMEREAGENT URICACIDSTANDARD(10mg/d1) SPECIMEN Useunhemolyzedserumorplasma. RecommendedanticoagulantsareEDTAorherapin. 0Uricacidserumorplasmaisreportedly(2)stablefor3daysat28oCandupto6 monthswhenfrozen(20oC). Serumuricacidisdestroyedwhenserumisallowedtoremainincontractwithredblood cellsandwhenserumiscontaminatedwithbacteria(2).

- 84 -

EZLabManual MANUALPROCEDURE Transfer2.5mlofURICACIDBUFFERtotestvialslabeled:UNKNOWN(S),STANDARD, CONTROL,BLANK.Add50microliters(ul)ofsampletoitsrespectivevial.Add100microlitersof URICACIDENZYMEREAGENTtoallvials,mixbygentleswirling,andallowtostandatroom temperature(<30oC)for10minutes.Zerophotometerat520+5nmusingBLANK.Measure absorbanceofallvialswithin15minutes. Note:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard.Readthe Unknownconcentrationdirectly. CALCULATIONWITHEXAMPLE: EXPECTEDVALUES(6) Children..2.05.5mg/d1 AdultMale...3.57.2mg/d1 AdultFemale2.66.0mg/d1 PROCEDURELIMITATIONS: Elevatedbilirubin(>9.94mg/d1)andascorbicacid(>190umol/Linserum)canresultin falseuricacidvalues(4).Thefollowingsubstancesreportedly(4)interfereinveryhighclinical doses:Methyldopa,Ldopa,3,4dihydroxyphenylaceticacid,gentisticacid. PROCEDURENOTES Uricacidexceedslinearity: Preparea1.1dilutionofthesamplewithReagentGradeWaterandrepeattest;multiply resultby2. Extremelylipemicsample: Add50microliterofsampleto2.6mlofReagentGradeWaterandmeasureabsorbance fromat520nmversuswaterinthesamesizecuvetasusedinabovePROCEDURE; subtracttheabsorbancefromtheoriginaltestabsorbanceandrecalculate: [(A(U)A(SB))/A(S)]xC(CS)=C(U) WhereSB=SampleBlank SIunitconversion: [(mg/d1)/16.9]=mmol/L

- 85 -

EZLabManual QUALITYCONTROL Goodlaboratorypracticerequirestheuseofnormalandabnormalcontrolseraatleast oncedailytomonitortestprecisionandaccuracy.WerecommendSterlingNormalControl(Cat. No.1003N)andSterlingAbnormalControl(Cat.No.1003A)forthispurpose. BLOODCHEMISTRY CREATININE(ModifiedHeinegardTiderstrom) INTENDEDUSE: FORINVITRODIAGNOSTICUSEinthequantitativecolorimetricendpointdeterminationof creatinineinserumandurine. REAGENTS: Thefollowingreagentscanbeuseduntiltheexpirationdateindicatedontheindividual bottlesandshouldbestoredatroomtemperature(30oC): CREATININEBUFFER: PICRICACID CREATININECLEARINGSOLUTION CREATININESTANDARD SPECIMEN SERUM Useunhemolyzedserum.Stableifrefrigerated(28oC)for24hours.Freezeforlonger storage. URINE Collect24hrurineincontainerwith15gmofboricacid.Stableforupto4daysif refrigerated(28oC).Freezeforlongerstorage. ENDPOINTPROCEDURE Transfer2.0mlofCreatinineBuffertovialslabeled: UNKNOWN(S),STANDARD,CONTROLandBLANK. Add1.0mlofPicricAcidReagenttoeachvialandmix Add200microlitersofsampletoitsrespectivevialandmix. (AddReagentGradeWatertoBlank). Note:ifassayingurine,usea1:100dilutionofurinewithReagentGradeWater. - 86 -

EZLabManual Placeallvialsin37oCheatingbathfor15minutes Zerothephotometerat520+5nmusingtheBLANK.Measureabsorbanceofsamples;theseare theA1readings. Add2dropsofCreatinineClearingSolutiontoeachvial,mixandletstandfor5minutesatroom temperature. Measureabsorbanceasbefore;thesearetheAoreadingsandshouldbemadewithin30 minutes. CALCULATIONWITHEXAMPLES SERUM WhereA=absorbance,U=UNKNOWN,S=STANDARD,C=concentration [U(A1Ao)/S(A1)]C(S)=C(U) (mg/d1)(mg/d1) EXAMPLE: U(A1)=0.12,U(Ao)=0.05,S(A1)=0.25,C(S)=5.0mg/d1,then: [(0.120.05)/0.25]x5.0mg/d1=1.4mg/d1 BLOODCHEMISTRY UREANITROGEN(BUN)(Berthelot/Colorimetric) INTENDEDUSE FORINVITRODIAGNOSTICUSEinthequantitativecolorimetricdeterminationofureanitrogen inserum REAGENTS Thefollowingreagentsshouldbestoredat28oCandcanbeuseduntiltheexpiration dateindicatedontheindividualbottle: UREANZYMEREAGENT UREANCOLORREAGENT UREANBASEREAGENT UREANSTANDARD(25mg/d1)

- 87 -

EZLabManual SPECIMEN Serum is recommended for the assay. Plasma may also be used, provided that anticoagulantusecontainsneitherammoniumnorfluoridesalts.Sodium,potassium,orlithium saltsofherapin,EDTA,oroxalatearesatisfactoryanticoagulants. Ureainserumisstableforupto24hoursatroomtemperature,forleastseveraldaysat o 46 C,andforatleast23monthswhenfrozen. PROCEDURE Transfer0.5mlofUREANCOLORRGTtovialslabeled: UNKOWN,CONTROL,STANDARD,BLANK. Add0.010ml(10ul)ofsampletoitscorrespondingvial. Add0.5mlofUREANZYMERGTtoallvials,mixgently,andincubateat37oCfor5 minutes*. Add2.0mlofUREANBASERGT,mixandincubateat37oCfor5minutes*. Setthewavelengthofthephotometerat630nmandzerothephotometerwiththe BLANK.ReadandrecordtheabsorbancesofallvialsandproceedtoCalculationwithExample below. Note:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard(25mg/d1). ReadtheUnknownconcentrationdirectly. *Alternative:Reactfor10minutesatroomtemperature226oC CALCULATIONWITHEXAMPLE WhereA=absorbance,U=UNKNOWN,S=STANDARD,C=concentration: A(U)xC(S)mg/d1=C(U)mg/d1 A(S) Example:A(U)=0.31,A(S)=0.48,C(S)=25mg/d1then: x25mg/d1=16mg/d1 0.48 EXPECTEDVALUES HealthyAmbulatoryAdults..718mg/d1 - 88 -

EZLabManual PROCEDURENOTES Ifasampleexceedsthelinearityofthetest,makeafivefolddilutionofsamplewithammonia freeReagentGradeWater(Cat.#10750)andrerunassay;multiplyresultby5. Toextendlinearityoftestto80mg/d1withoutdilutionofsample,doublereagentvolumeto 1.0mlUreaNColorRgt.,1.0mlUreaNZyme,4mlUreaNBaseReagentandretainthesame samplesizeof10ul. QUALITYCONTROL Qualitycontrolserashouldbeusedroutinelytomonitortestprecision(7).Refertothe manufacturers package insert for analyte stability and acceptable limits. Sterling Normal Control (Cat. # 1003N) and Sterling Abnormal Control (Cat. # 1003A) are recommended for thispurpose. BLOODCHEMISTRY ENZYMATICGLUCOSE(Trinder/GODPOD) Collectwholebloodybyvenipunctureandallowtoclot.Contrifugeandremoveserum assoonaspossible.Bloodshouldbecollectedafter1216hoursfastingtominimizeturbidity; however, the effect of a meal on the cholesterol level reportedly (8) increases the level less than3%. Cholesterolreportedly(9)decreasesduringovulation,increasesduringseveremotional stress, varies in some individuals on a day today basis, and exhibits a seasonal variation in healthyyoungmen(9). MANUALPROCEDURE All reagents must attain room temperature (2025oC) prior to use. Transfer 2.5 ml of cholesterol Buffer to vials labeled: UNKNOWN (S), STANDARD, CONTROL, BLANK. Add 20 ul (0.020ml)ofsampletoitsrespectivevial.Add0.1ml(100ul)ofChloresterolEnzymeReagentto allvials,mixandincubateat37oheatingbathfor10minutes. Zerophotometerat510nmwiththeBLANKvial.Readandrecordtheabsorbanceofall vials.
Note:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard(200mg/d1).Read theUnknownconcentrationdirectly.

UsetheabsorbancereadingsoftheCholesterolSTANDARDandUNKNOWN(S) tocalculatethetotalcholesterolvaluesasfollows: A(U)xC(S)mg/d1=C(U)mg/d1 A(S) - 89 -

EZLabManual EXPECTEDVALUES AGE(YRS) MALES(mg/d1)FEMALES(mg/d1) 519 125191 131208 2034 118258 121227 ` 3549 147275 139268 5064 156287 163300 6570+ 166265 167280 DESIREDRANGEFORADULTS(10) 140250mg/d1 PROCEDURENOTES Ifthetotalcholesterolvalueexceeds600mg/d1orthelinearcapabilityofthephotometer, dilutethesample1:1withdistilledwaterandreruntheassay;multiplytheresultby2. Ifthesampleisextremelylipemic,prepareasampleblankbyadding0.020ml(20ul)ofsample to2.5mlCholesterolBufferandadd0.1mlofdistilledwater.Mixandmeasuretheabsorbance againstCholesterolBuffer.Subtractthevalueobtainedfromthevaluederivedinstep#4ofthe aboveprocedure;thiswillyieldacorrectedabsorbancevalue. QUALITYCONTROL GoodLaboratorypracticerequirestheuseofnormalandabnormalcontrolseraatleast oncedailytomonitortestprecisionandaccuracy.WerecommendSterlingNormalControl(Cat. #1003N)andSterlingAbnormalControl(Cat.#1003A)forthispurpose. BLOODCHEMISTRY TRIGLYCERIDES(Colometric) INTENDEDUSE FORINVITRODIAGNOSTICUSEinthequantitativecolormetrideterminationofserum triglycerides. REAGENTS TRIGGPOBUFFER TRIGGPOENZYMEREAGENT TRIGLYCERIDESTANDARD SPECIMEN - 90 -

EZLabManual Collect whole blood from fasting patients, allow to clot, centrifugeand revome serum with2hoursofdrawing.Collectiontubesmustnotcontainglycerollubricant. MANUALPROCEDURE All reagents must attain room temperature (2025 oC) prior to use. Transfer 1.0 ml of WORKINGTRIGGPOtovailslabeled:UNKNOWN,CONTROL,STANDARD,REAGENTBLANK. Add0.010 ml(10ul)ofsampletoitsrespectivevial,mixbygentleswirlingandplacein37oC heatingbathfor10minutes.Removevialsfromheatingbathandadd1.5mlofReagentGrade Watertoallvailsmixbygentleswirling. Note: The addition of 1.5 ml of Reagent Grade Water is necessary only if your photometer requiresafinalvolumeinexcessof1.0ml;otherwise,youmaymeasurethefinalcolordirectly. ZerothephotometerwiththeREAGENTBLANKat515+5nm.Recordabsorbanceofallvialsand proceedtoCalculationWithExamplebelow. Note:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard(250mg/d1). ReadtheUnknownconcentrationdirectly. PROCEDURELIMITATIONS: Grosslyictericsamples(>10mg/d1bilirubin)shouldnotbeused. EXPECTEDVALUES Adults: 10190mg/d1 PROCEDURENOTES For grossly icteric or lipemic samples, prepare a SAMPLE BLANK by adding 0.01. ml of sampleto2.5mlof0.85%salineandmeasureitsabsorbanceversus0.85%salineat515nm. SubtractthisabsorbancefromtheTESTabsorbanceobtainedinthePROCEDURE. ForlaboratoriesusingtheSIsystem: Mg/d1x0.0113=mmol/L (Expressingtriglyceridesastriolein) QUALITYCONTROL Qualitycontrolserashouldbeusedroutinelytomonitortestprecision(16).Refertothe manufacturerspackageinsertforanalytestabilityandacceptablelimits.SterlingNormal - 91 -

EZLabManual Control(Cat.No.1003N)andSterlingAbnormalControl(Cat.No.1003A)arerecommended forthispurpose. BLOODCHEMISTRY HDLCHOLESTEROL(PEG6000method) INTENDEDUSE FORINVITRODIAGNOSTICUSEinthequantitativedeterminationofhighdensitylipoprotein cholesterol(HDLC)inserum. REAGENTS Thefollowingreagentsshouldbestoredatroomtemperature(<3oC)andcanbeused untiltheexpirationdateindictedontheindividualbottle: HDLPEGREAGENT HDLCHOLESTEROLSTANDARD SPECIMEN Collectwholebloodfromafastingpatientbyvenipunctureandallowtoclot.Centrifuge andremoveserum.HDLcholesterolisreportedlystableforupto1weekat4oC(5)or20oC(8). Hemolyzedspecimenshouldnotbeusedforthisassay. PRECIPITATIONSTEP: Transfer200microlitersoftheHDLPEGREAGENTtoamicrocentrifugetubelabeled: UNKNOWN(S),CONTROL,etc.Add200microlitersofsampletoitsrespectivemicrocentrifuge tube,capandmixvigorously.Usinga13x100mmtesttubeasacarrier,centrifugeat1000 2000XGatleast10minutesProceedwithCholesterolassay. CHOLESTEROLASSAY All reagents must attain room temperature (2025oC) prior to use. Add 2.5 ml of CholesterolBuffertovialslabeled:UNKNOWN(S),STANDARD,CONTROL,BLANK.Add100ulof the supernate (from Precipitation Step) to its respective vial. Add 100 ul of HDL Cholesterol StandardtotheSTANDARDvial.Mixallvials.Add100ulofCholesterolEnzymeReagenttoall vials,mixandreturntothe37oCheatingbathfor10minutes.Zerophotometerat510nmwith theBLANKvial.Readandrecordtheabsorbanceofallvials. Note:Foradirectreadoutinstrument,setreadoutto100mg/d1(correctedforthedilution factor)andreadtheUNKNOWNconcentrationdirectly. - 92 -

EZLabManual EXPECTEDVALUES HDLCHOLESTEROL(mg/d1) MaleFemale CordBlood 550 550 014yr 3065 3065 510yr 3065 3065 2029yr 3070 3070 3039yr 3070 3080 40+yr 3070 3085 Valuesforblanksareapproximately10mg/d1higherthanthemeansandrangesgivenabove. PROCEDURENOTES Centrifugationshouldaclearsupernatant.Sinceacloudysupernatantwillyieldfalselyelevated values,thesampleshouldbecentrifugedagaintogiveaclearsupernate. Dilutionofalipemicspecimenwithsalineisnotrecommendedowingtoreportedlylarge dilutionerror(10).Ifadilutionmustbeperformed,a5%w/vsolutionofBovinealbuminin100 mmol/LaceratebufferatpH7.5+0.5,isrecommendedasthediluent. LDLCholesterolCalculation LDLCholesterol=TotalCholesterolHDLCholesterol*VLDLChol.*VLDLCholesterol= (Triglyceride) 4.6 QUALITYCONTROL Qualitycontrolserashouldbeusedroutinelytomonitortestprecision(7).Refertothe manufacturerspackageinsertforanalytestabilityandacceptablelimits.SterlingNormal Control(Cat.#1003N)andsterlingAbnormalControl(Cat.#1002A)arerecommended. URINE WhereA=absorbance,U=UNKNOWN,S=STANDARD,C=concentration,0.01=factorforurine dilution,V=volumeofurinefor24hours(liters): [U(A1Ao)/S(A1)]xC(S)x0.01x100V=C(U) EXAMPLE: U(A1)=0.09,U(Ao)=0.02,S(A1)=0.25,C(S)=5.0mg/d1,V=1700ml/24hours=1.7L/24hrs, then: [(0.090.02)/0.25]x5.0mg/d1x0.01x100x1.7L/24Hrs=2.38Gm/24Hrs - 93 -

EZLabManual CREATININECLEARANCE [UrineCreatinine(mg/d1)xUrineVol(ml/min)]/SerumCreatinine(mg/d1)=Creatinine Clearance(ml/min) Usingabovecalculationsandconverting1700mlurine/24hrsto1.18ml/min,then: [(140mg/d1x1.18ml/min)/1.4mg/d1)]=118ml/min EXPECTEDVALUES(6) MALESFEMALES SERUM(mg/d1) 0.71.30.61.2 URINE(Gm/24hrs) 0.82.00.61.8 CREAT.CLEAR(ml/min) 97.137 88.128 PROCEDURENOTES The Ao absorbance reading of the STANDARD should be 0.00 since the addition of acid is designedtolowerthepHanddestroyallofthealkalinecreatininepicratecomplex. Ifthecreatininevalueexceeds15.0mg/d1,repeattheassayon1:1dilution;multiplyresultby 2. Ifturbidityoccursintestvials(step#6inEndpointProcedure)aftertheadditionof.1mlof CreatinineSolution,werecommendeitherofthefollowingoptions: Centrifuge vials at high speed (20003000 RPM) and read absorbance over the small protein buttoninthebottomofvial. Repeattestusing0.05ml(1drop)ofCreatinineClearingSolutioninsteadofthespecified0.1 ml. QUALITYCONTROL Goodlaboratorypracticerequirestheuseofnormalandabnormalcontrolseraatleast oncedailytomonitortestprecisionandaccuracy.WerecommendSterlingNormalControl(Cat. No.1003N)andSterlingAbnormalControl(Cat.No.1003A)forthispurpose.

- 94 -

EZLabManual BLOODCHEMISTRY OGTTANDGCT

OGTTFORNONPREGNANTADULT 1. ExtractbloodforFBS 2. Askpatienttodrink75gramsglucose 3. Getserumglucose2hoursafterdrinking. SCREENINGTESTFORPREGNANTPATIENT(GCT) 1. Askpatienttodrink50gramsglucose(2428wks) 2. Getserumglucose1hourafterdrinking. DIAGNOSTICTESTFORPREGNANTPATIENTS(OGTT) 1.ExtractbloodforFBS. 2.Askpatienttodrink100gramsglucose. 3.Getserumglucose1hourafterdrinking. 4.Getserumglucose2hoursafterdrinking. 5.Getserumglucose3hoursafterdrinking. BLOODCHEMISTRY POTASSIUMREAGENTSET(TURBIDIMETRIC) INTENDEDUSE FOR IN VITRO DIANOSTIC USE in the quantitative determination of potassium in serum and urine. REAGENTS 1. PROCIPREAGENT 2. BORONREAGENT 3. BASEREAGENT 4. POTASSIUMREAGENT SPECIMENCOLLECTION SERIUM Collect whole blood by venipuncture and allow to clot. Centrifuge and immediately removeserum.Serumshouldnotbeallowedtoremainonclot.Prolongedvenousstatisduring venipuncturereportedly(6,7)causes1020%increaseinthelevel.SincetheconcentrationofK - 95 -

EZLabManual inerythrocytesisabout20timesthatinserumorplasma(8),itsimperativethathemolysisbe avoided. PLASMA Hemolysisfree plasma may also be used. It must also be rapidly separated from the cells.Ofcourse,anticoagulantscontainingpotassiummustnotbeused.Heparinizedplasmais preferred. MANUALENDPOINTPROCEDURE A.DEPROTEINIZATION 1.Labelpolyethylenemicrocentrifugetubes:UNKOWN(S),STANDARDandCONTROL. 2.Transfer1.0ml(100ul)ofsampletootsrespectivevial. 3.Add1.0mlofProCipReagent,capandmixvigorously.Letstandforatleast3minutes. 4.Usinga13x100mmtesttubeasacarrier,centrifugeat2000xg(25003000rpm)for5 minutes.Testisperformedonthesupernatantfluid. B.REACTION 1.Labeltestvials:UNKOWN(S),STANDARD,CONTROL,andREAGENTBLANK. 2.ADD2.0MLOFBoronreagenttoallvials. 3.Add2.0ml(4drops)ofbaseReagenttoeachvialandmiximmediately. 4.Add0.5mlNote:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard (5.0mEq/L).readtheunknownconcentrationdirectly. EXPECTEDVALUES(6) (500ul)ofSupernatantfluidfromabovetoitsrespectivevialandmixbygentleswirlingto avoidfoaming.Add0.5mlofReagentGradeWatertotheREAGENTBLANKandmix. 5.Setwavelengthofphotometerat420nmandzerotheinstrumentwiththeREAGENTBLANK. Readandrecordtheabsorbanceofallvials. Serum 3.65.5mEq/l Plasma 3.44.5mEq/L Urine 25125mEq/L (Varieswithdiet)
+

- 96 -

EZLabManual BLOODCHEMISTRY SODIUMREAGENTSET(ColorimetricUranylReductionMethod) INTENDEDUSE FOR IN VITRO DIANOSTIC USE in the quantitative colorimetric determination of sodium in serumandplasma. REAGENT 1. SODIUMPROCIPREAGENT 2. SODIUMACIDREAGENT 3. SODIUMCOLORREAGENT PROCEDURE A.DEPROTEINIZATION 1.Labelpolyethylenemicrocentrifugetubes:UNKNOWN(S),STANDARD,CONTROL,andBLANK. 2.Add1.0mlofSodiumProcipReagenttoeachvial. 3.Transfer20microlitersofsampletoitsvial,capandmixvigorously.Letstandfor10minutes inanicebath(00c). 4.Mixtubesvigorously.Usinga13x100mmtesttubeacarrier,centrifugeat2000g(25003000 rpm)for10minutes. B.COLORREACTION 1.Labelborosilicatetestvials:UKNOWN(S),STANDARD,CONTROLBLANK. 2.Add1.5mlofSodiumAcidReagenttoallvials. 3.Carefullytransfer50microlitersofsupernatantfromDEPROTEINIZATIONstepabovetoits respectivevial. 4.Add1.5mlofSodiumColorReagenttoeachvialandmixbygentleswirling. 5.Setthewavelengthofthephotometerat550+10nmandzerotheinstrumentwithdistilled water.Readandrecordtheabsorbancesofallvialswithin60minutes. EXPECTEDVALUES(7) Adults.. 135155mEq/L Thereappearstobesignificantsexdifference.Thereisaslightincreasingaftermeals.

- 97 -

EZLabManual BLOODCHEMISTRY TOTALCHOLESTEROL(ENZYMATICMETHOD)

INTENDEDUSE FORINVITRODIAGNOSTICUSEinthequantitativedeterminationoftotalcholesterolinserum. REAGENTS 1. CHOLESTEROLBUFFER 2. CHOLESTEROLENZYMEREAGENT 3. CHOLESTEROLSTANDARD SPECIMEN Collectwholebloodybyvenipunctureandallowtoclot.Contrifugeandremoveserum assoonaspossible.Bloodshouldbecollectedafter1216hoursfastingtominimizeturbidity; however, the effect of a meal on the cholesterol level reportedly (8) increases the level less than3%. Cholesterolreportedly(9)decreasesduringovulation,increasesduringseveremotional stress, varies in some individuals on a day today basis, and exhibits a seasonal variation in healthyyoungmen(9). MANUALPROCEDURE Allreagentsmustattainroomtemperature(2025oC)priortouse. 1. Transfer 2.5 ml of cholesterol Buffer to vials labeled: UNKNOWN (S), STANDARD, CONTROL,BLANK. 2. Add20ul(0.020ml)ofsampletoitsrespectivevial 3. Add0.1ml(100ul)ofChloresterolEnzymeReagenttoallvials,mixandincubateat 37oheatingbathfor10minutes. 4. Zerophotometerat510nmwiththeBLANKvial.Readandrecordtheabsorbanceof allvials. Note:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard (200mg/d1).ReadtheUnknownconcentrationdirectly.

- 98 -

EZLabManual Use the absorbance readings of the Cholesterol STANDARD and UNKNOWN (S) to calculatethetotalcholesterolvaluesasfollows: A(U)xC(S)mg/d1=C(U)mg/d1 A(S) EXPECTEDVALUES AGE(YRS) MALES(mg/d1)FEMALES(mg/d1) 519 125191 131208 2034 118258 121227 ` 3549 147275 139268 5064 156287 163300 6570+ 166265 167280 DESIREDRANGEFORADULTS(10) 140250mg/d1 PROCEDURENOTES 1. If the total cholesterol value exceeds 600 mg/d1 or the linear capability of the photometer,dilutethesample1:1withdistilledwaterandreruntheassay;multiply theresultby2. 2. Ifthesampleisextremelylipemic,prepareasampleblankbyadding0.020ml(20ul) of sample to 2.5 ml Cholesterol Buffer and add 0.1 ml of distilled water. Mix and measure the absorbance against Cholesterol Buffer. Subtract the value obtained fromthevaluederivedinstep#4oftheaboveprocedure;thiswillyieldacorrected absorbancevalue. QUALITYCONTROL GoodLaboratorypracticerequirestheuseofnormalandabnormalcontrolseraatleast oncedailytomonitortestprecisionandaccuracy.WerecommendSterlingNormalControl(Cat. #1003N)andSterlingAbnormalControl(Cat.#1003A)forthispurpose.

- 99 -

EZLabManual BLOODCHEMISTRY CALCIUM(CPC)PROCEDURE INTENDEDUSE FORINVITRODIAGNOSISUSEinthequantitativecolorimetricdeterminationoftotalcalciumin serum. REAGENTS 1.CALCIUMCOLORREAGENT 2.CALCIUMBASEREAGENT 3.CALCIUMSTANDARD SPECIMENCOLLECTION&HANDLING Collect whole blood by venipuncture and allow to clot. Centrifuge and immediately remove serum. The specimen should be collected from a fasting individual since meals reportedly (9) increase calcium levels as a result of metabolic alkalosis. Serum must be used mostanticoagulantsinterferewithcomplexometricmethods. Once separated from the cells, calcium in serum is stable for 8 hours at room temperature,24hoursat40C,and1yearfrozen(10). MANUALPROCEDURE 1. Transfer 1.5ml of CALCIUM COLORREAGENT into vials labeled: UNKNOWN(S), STANDARD,BLANK,etc. 2. Add1.5mlofCALCIUMBASEREAGENTtoallvialsandmixbygentleswirling 3. Add20ul(0.020ml)ofsampletoitsrespectivevial,mixbygentleswirlingandallowto standatroomtemperature(20250C)foratleastone(1)minute. 4. Set the wavelength of photometerat 575+5 nm and zeroinstrumentwith the BLANK. Readandrecordtheabsorbancesofallvials. Note: For a direct readout instrument, set readout to concentration of Standard (10mg/dl).ReadtheUnknownconcentrationdirectly. EXPECTEDVALUES Newborn(47days) 9.810.9mg/dl Children 8.810.8mg/dl Adult(M&F) 8.410.2mg/dl AdultM>60yrs 8.410.0mg/dl

- 100 -

EZLabManual PROCEDURENOTES 1.PreparationofSERUMBLANKforhighlylipemicsamples: a.Add20ul(0.20ml)ofthesampleto3.0mlofdistilledwaterandmix. b.ReadtheabsorbanceofthisSERUMBLANKagainstdistilledwaterat575+5nmand subtractthisabsorbanceoftheUKNOWN(fromstep#4oftheaboveprocedure). c.UsethecorrectedabsorbancetocalculatethecalciumvalueintheUNKNOWN. 2. If any assay value exceeds 15 mg/dl or the linear capability of the photometer, repeat the testona1:1dilutionofthesamplewithdistilledwaterandmultiplytheresultby2. 3.Contaminationoftheglasswarewithcalcium,usuallyfromdetergents,maybepreventedby usingacidwashedglassware. QUALITYCONTROL Goodlaboratorypracticerequirestheuseofnormalandabnormalcontrolseraatleast oncedailytomonitortestprecisionandaccuracy.WerecommendedSterlingNormalControl (Cat.#1003N)andSterlingAbnormalControl(Cat#1003A)forthispurpose. BLOODCHEMISTRY ALBUMINBCGPROCEDURE INTENDEDUSE FORINVITRODIAGNOSISUSEinthequantitativedeterminationofSerumAlbumin. REAGENTS 1. ALBUMINBCGREAGENT 2. ALBUMINSTANDARD SPECIMENCOLLECTION(6) Collect whole blood, allow to clot, centrifuge and remove serum. Albumin in serum is reportedtobestableforupto7daysatroomtemperature,30daysat280C,andlongerwhen frozen. MANUALPROCEDURE 1. Place3.0mlofALBUMINBCGREAGENTintotestvialslabeled:UNKNOWN, CONTROL,STANDARD,BLANK. 2. Transfer0.010ml(10ul)ofsampletothecorrespondingvial,mixthoroughly, andallowtostandatroomtemperaturefor5minutes. - 101 -

EZLabManual 3. Setwavelengthofthephotometerat630nmandzerothephotometerwith theBLANK.Readtheremainingvialsandrecordtheabsorbances.

ALTERNATIVEMETHOD:Theabsorbancemaybereadat30seconds. EXPECTEDVALUES(7) New: 2.84.4Gm/dl (24days) Adult: 3.45.0Gm/dl >60yrs: 3.44.8Gm/dl PROCEDURENOTES 1. Albumin levels exceeding 6.0 Gm/dl must be diluted and reassayed. Dilute the sample1:1with0.85%saline,reruntestandmultiplyresultby2. 2. SERUMBLANKSmustberunwithmarkedlylipemicsample.Simplysubstitute3.0mlof distilled water for Albumin BCG Reagent, add 0.010 (10ul) of sample and take absorbancemeasurementat630nmversusdistilledwater.Subtractthisvaluefromthe absorbancevalueofinitialreadingofthelipemicspecimenintheaboveprocedure. QUALITYCONTROL Quality control sera should be used routinely to monitor test precision (11). Refer to the manufacturesproductinsertforanalytestabilityandacceptablelimits.SterlingNormalControl (CAT.#10024)arerecommendedforthispurpose. BLOODCHEMISTRY TOTALPROTEIN INTENDEDUSE FORINVITRODIAGNOSISUSEinthequantitativecolorimetricdeterminationofSerumTotal Protein. REAGENTS 1. BIURETREAGENT 2. TOTALPROTEINSTANDARD

- 102 -

EZLabManual SPECIMEN(4) Collect whole blood, allow to clot, centrifuge and remove serum. Protein in serum is reportedtobestableforupto7daysatroomtemperature. MANUALPROCEDURE 1. Place3.0mlofBiuretReagentintotestvialslabeled:UNKNOWN,CONTROL,STANDARD, BLANK. 2. Transfer 0.050 ml (50ul) of sample to the corresponding vial, mix thoroughly (but not vigorously),andallowtostandatroomtemperaturefor10minutes. 3. Set the wavelength of the photometer at 540 nm and zero the photometer with the BLANK.Readtheremainingvials,recordtheabsorbanceandproceedtocalculationwith Example. NOTE:Foradirectreadoutinstrument,setreadouttoconcentrationofStandard.Readthe Unknownconcentrationdirectly. EXPECTEDVALUES(6) 6.38.3Gm/dl PROCEDURENOTES Fortotalproteinlevelsexceeding15.0Gm/dl,reruntheprocedureona1:1dilutionofsample with0.85%salineandmultiplyresultby2. BLOODCHEMISTRY LACTATEDEHYDROGENASE(LCH)(COLORIMETRICMETHOD) INTENDEDUSE: FORINVITRODIANOSISUSEinthequantitativecolorimetricdeterminationoflactate dehydrogenase(LDH)inserum. REAGENTS Thefollowingreagentsmustbestoredunderrefrigeration(280C)andcanbeuseduntil theexpirationdateindicatedontheindividuallabel. 1. LDHSUBSTRATE 2. LDHCOLORREAGENT 3. LDHCALIBRATOR - 103 -

EZLabManual PROCEDURE Allreagentsmustattainroomtemperature(20250C)priortouse. Label vials: UNKNOWN(S), CONTROL, LDH CALIBRATOR and BLANK. Add 20 microliters ofsampletoitsrespectivevials. Add1.0mlofLDHSubstrateallvials. Add 100 microliters of LDH Color Reagent to all vials, mix and place in a 370C heating bathforexactly10minutes. Add2.0mlof0.1NHydrocholoricAcidtoeachvialtostopthereaction.Mixbygentle swirling. Zero the photometer at 500nm with the Reagent Blank. Read and record the absorbanceofallsampleswithin60minutes.

1. 2. 3. 4. 5.

EXPECTEDVALUES(12) 0 2777/L(37 Cor81231mU/ml(wackerunits;370C) PROCEDURENOTES SampleBlankPreparationforhighlyictericorlipemicspecimen: a. Add20microliterofsampleto3.1mlofreagentGradeWaterandmix. b. Zerophotometersetat500nmwithReagentGradeWaterandrecordsamplereading. c. Subtractthisvaluefromtheoriginalabsorbancevalueobtainedonthepatientsample andrecalculatetheLDHvalue. QUALITYCONTROL Good laboratory practice requiresthe use of normal and abnormal control sera at least once dailytomonitortestprecisionandaccuracy.WerecommendSterlingAbnormalControls(CAT# 1003N)andSterlingAbnormalControl(CAT#1003A)forthispurpose. BLOODCHEMISTRY ALKALINEPHOSPHATASE INTENDEDUSE FortheQuantitativeDeterminationofSerumAlkalinePhosphatase. REAGENTS Thefollowingreagentsshouldbestoredat280Candcanbeuseduntiltheexpirationdate indicatedontheindividualbottles. - 104 -

EZLabManual REAGENTS 1. 2. 3. 4. SPECIMENCOLLECTION(3,4) Collectwholebloodbyvenipunctureandallowittoclot.Centrifugeandremoveserum. Hemolysis must be avoided since ALP in erythrocytesis six times the concentration found in normal serum. Serum must be used for this assay because most anticoagulants nhibit ALP activity. Serumshouldbeseparatedfromthecellswithin2hoursfollowingcollectionand,ifnot assayedpromptly,serumshouldberefrigeratedat280C.Ifstorageistoexceed23days,serum shouldbefrozenandstoredat200C.PleasenotethatALPactivityrecoversslowlyafterthawing offrozenserum. PROCEDURE Allreagentsmustattainroomtemperature(20250C)priortouse. 1. Add 1 drop of ALP Activity to all vials labeled:UNKNOWN (S), CONTROL, STANDARD and BLANK. 2.Add50ulofsampletoitsrespectivevial. 3.Add1.0mlofALPSubstratetoallvials,mixbyshakingentirerack,andimmediatelyplacein 370Cheatingbathforexactly10minutes. 4.Add2.0mlofALPDILUENTtoallvialsandmix. 5.Zerophotometerat590nmwithReagentGradeWater. Note:ForaSA2000ChemistryAnalyzer,readSTANDARDandsampleagainstBLANK. EXPECTEDVALUES(8) 945AT370C PROCEDURENOTES 1.Ifthesampleislipemic,highlyictericorcontainsBSP,aSERUMBLANKshouldbeincludedas follows: a.Add1.0mlofALPSUBSTRATE,1dropofALPACTIVATORand2.0mlofALPDILUENTto atestvialthatisidenticaltothatusedinaboveMANUALPROCEDURE. b.Transfer0.050ml(50ul)ofsampletovialandmixbygentleswirling. c.ReadandrecordtheabsorbanceoftheSERUMBLANKvs.theReagentBLANKinSA 2000ChemAnalyzer. - 105 ALPSUBSTRATE ALPACTIVATOR ALPDILUENT ALPSTANDARD(50IU/L)

EZLabManual d.Calculateasfollow:[a(Serum)a(SerumBlank)]/a(Standard)XConcentrationof Standard(IU/L)=ALPConcentrationinserum(IU/L) BLOODCHEMISTRY TROPONINTSENSITIVERAPIDASSAY PROCEDURE: 1. Removethestripfromthepackageandplaceitonaflatsurface. 2. ObtainthebloodsamplecollectedatEDTAtubes. 3. Drawup150ulbloodintothedispensingaid. 4. Applythesampletotheteststripandreadresultafter15minutes. INTERPRETATIONOFRESULTS: NEGATIVEonlysinglelineatcontrolline POSITIVEtwolinesappearatthetestandcontrolline BLOODCHEMISTRY TROPONINI SPECIMENCOLLECTIONANDSTORAGE: 1.Serum 2.Wholeblood.Collectbloodinatubecontainingcitrateastheanticoagulant.Freshspecimen arerecommendedsincethecardiactroponinproteinsareunstable.Wholebloodsampleshould betestedwithinfourhours. PROCEDURE: 1. Refrigeratedspecimenorothertestmaterialsincludingdevicesmustbeequilibratedto roomtemperaturebeforetestingtoavoidinvalidresults. 2. Removethedevicefromthepouchandplaceitonaflatsurface.Labelthedevicewith specimenidentification. 3. Addfourdropsofwholebloodorserum(about160ul200ul}intothesamplewell. 4. Strongpositiveresultsmaybeobservedwithinfiveminutes.Weakpositiveresultsmay takealongertime.THERESULTSSHOULDBEREADWITHIN1520MINUTES.DONOT INTERPRETTHERESULTSAFTER20MINUTES. INTERPRETATIONOFRESULTS: 1. POSITIVE If both the C line and T line appear, the results indicates that the cardiac troponinisdetectedandtheresultispositive. - 106 -

EZLabManual 2. 3. NEGATIVE. If only the C line appears, the test indicates that no cardiac troponin is detectedoritslevelisbelowthedetectable. INVALIDWhennocontrollineappearswithin5minutes.Repeatthetestwithanew testdevice.

BLOODCHEMISTRY CREATINEKINASEMB(CKMB) INTENDEDUSE CKMBisintendedtomeasuretheactivityofisoenzymeCKMBinhumanserumat340nm. PROCEDURE(MANUAL) 1. ReconstituteCKMBreagentaccordingtoinstructions. 2. Pipette 1.0 ml of CKMB reagent into the appreciate test tubes and prewarm at 37oCforleasttwo(2)minutes. 3. Zerospectrophotometerwithwaterat340nm. 4. Add0.050ml(50ul)ofsampletothereagent,mix,andincubateat37oCforfive(5) minutes. 5. Afterfiveminutes,readandrecordthechangeinabsorbanceperminutefortwo(2) minutes 6. Calculatetheaverageabsorbancedifferenceperminute(Abs./min.). 7. TheAbs./min.Multipliedbythefactor3376(seecalculations)willyieldCKBresults inIU/L. 8. Sample with values above 1500 IU/L should be diluted 1:1 with saline, re assayed, andtheresultmultipliedbytwo(2). Note:Ifthespectrophotometerbeingusedrequiresafinalvolumegreaterthan1.0mlfor accuratereadings,3.0mlofreagentand0.15ml(150ul)ofsamplecanbeused.Ifthe spectrophotometerbeingusedisequippedwithatemperaturecontrolledcuvette,thereaction mixturemaybeleftinthecuvettewhilereadingsaretaken. EXPECTEDVALUES 024IU/L(37oC)

- 107 -

EZLabManual BLOODCHEMISTRY PHOSPHORUSUV Samples Serumfreeofhemolysis Heparinplasma Urinedituled1/10withdistilledwater ReferencesValues Serum Urine Procedure Wavelength :340nm Temperature :250C,300C,370C Cuvette :1cmlightpath Readagainstreagentblank BLANK WorkingReagent 1ml Distilledwater 10ul Standard Sample

2.74.5 2745 0.871.45 325900 10.428.8

mg/dL mg/L mmol/L mg/24h mmol/24h

STANDARD 1ml 10ul

SAMPLE 1ml 10ul

Mixandreadtheopticaldensity(OD)aftera5minutesincubation. Thefinalcolourisstableforatleast1hour. - 108 -

EZLabManual SPECIALTEST HEPATITISBSURFACEANTIBODY SPECIMENCOLLECTIONANDSTORAGE: Collectbloodasepticallybyvenipunctureintoacleantubewithoutanticoagulants. Permitbloodtoclotfortwentytothirtyminutesatroomtemperature.Centrifuge Toobtainclearserumandtransferserumintoacleanglasstube.Thetestmaybe Performedusinghumanserumorplasma. TESTPROCEDURE: NOTE:Bringtestcomponentsandspecimenstoroomtemperaturepriortotesting. 1.Openafoilpouch,removetheTestingDeviceandplaceitonaclean,level surface. 2.Holdingthedropperprovidedvertically,addexactlyfourdropsofserumtothe samplewellS. 3.Readthetestresultattenminutes. IMPORTANT:APOSITIVERESULTshouldnotchangeonceestablishedat10minutes;however, inordertopreventanincorrectreading,donotinterpretthetest resultaftermorethantenminutes. INTERPRETATIONOFRESULTS: 1.POSITIVE.Twopinkrosebandsappear;oneinthetestzoneTandinthecontrolzoneC.A positiveresultindicatesthepresenceoftheantibodytohepatitisBantigen. 2.NEGATIVE.OnepinkrosebandappearsinthecontrolzoneC,withabandinthetestzone T.AnegativeresultindicatestheantibodytohepatitisBantigenisnotpresent,oritslevelis belowthesensitivityof thetest. 3.INVALID.Ifthereisnodistinctcolorbandvisiblebothinthetestzoneandinthecontrol zone,orthereisavisiblebandonlyinthetestzoneandnotinthecontrolzone,thenthetestis INVALID.

- 109 -

EZLabManual SPECIALTEST

HEPATITISBSURFACEANTIGENScreeningTest

SPECIMENCOLLECTIONANDSTORAGE: 1.Serumisobtainedbyfollowingregularclinicalprocedures.Ifthespecimenisnottestedthe same day as collection, it should be sealed and refrigerated. If not tested with in 3 days of collection ,the specimen should be frozen. Avoid thawing and refreezing of the specimen. Sodiumazidecanbeaddedasapreservationupto0.1%withoutanyeffectontheassay. 2.Ifspecimensaretobeshipped,theyshouldbepackedincompliancewithfederalregulations coveringthetransportationofetiologicagents. 3.Specimenscontainingprecipitatemayyieldinconsistenttestresults. Suchspecimensmustbeclarifiedpriortoassaying. PROCEDURE: 1.Bringallreagentsandsamplestoroomtemperature. 2.Removethetestdevicefromitsfoilpouch. 3.Dispense200ulabout4dropsofserumsampleintothesamplewellwithacleanpipette. 4.Readtheresultwithin10minutesfor5ng/ml,orwithin30minutesfor1ng/ml. Note:Donotinterprettheresultafter330minutes. INTERPRETATIONOFRESULTS: 1.NEGATIVE:Onlyonecoloredbandappearsonthecontrolregion. 2. POSITIVE: In addition to thecontrol band, adistinct colored band also appearson the test region. SPECIALTEST SDDENGUEIgG/IgM PROCEDUREOFTHETEST 1.Allowallkitcomponentsandspecimentoroomtemperaturepriorto testing. 2.Add4drops(about140ul)ofassaybuffertoeachdisposabletest tube. 3.Usingtheloopsprovided(*),add1ulofspecimentotesttube containingassaybuffer. (*Dipthecircularendofthespecimenloopintothespecimen,andthen carefullyplacethecircularendoftheloopintothetesttube.Thiswilladd 1ulofspecimentoassaybuffer.) - 110 -

EZLabManual 4.Gentlystirtheassaybufferwiththelooptoensureadequatemixingof thespecimenintheassaybufferandremovetheloop. 5.Removetheteststripfromthefoilpouchpriortouse. 6.Holdingthestripvertically,inserttheteststripintothetubecontaining 7.Interprettestresultat1530minutes.Ifthetestbandisveryfaintat15 minutes,pleasereadtheresultsagainat30minutes. INTERPRETATIONOFTHETEST IgGandIgMpositive Thecontrolline,IgM(M)andIgG(G)arevisibleontheteststrip.ThisispositiveforbothIgM andIgGantibodies.Thisisindicativeoflateprimaryorearlysecondarydengueinfection. IgMPositive The control line (C) and IgM line (M) are visible on the est strip. This is positive for IgM antibodiestoDenguevirus.ThisisindicativeofaprimaryDengueinfection. IgGpositive The control line, IgG (G) are visible on the test strip. This is positive IgG antibodies.This is indicativeofsecondaryorpastdengueinfection. Negative Thecontrollineisonlyvisibleontheteststrip.NoIgGandIgMantibodiesweredetected.Retest in35daysifdengueinfectionissuspected. SPECIALTEST SDSALMONELLATYPHIIGG/IGM Procedureofthetest
Allowallkitcomponentsandspecimentoroomtemperaturepriorto testing. Addfourdropsabout20ulassaybuffertoeachdisposabletesttube. Usingtheloopsprovidedadd1ulofspecimentotesttubecontaining assaybuffer.DIPthecircularendofthespecimenloopintothe

specimen,andthencarefullyplacethecircularendoftheloopinto thetesttube.Thiswilladd1ulofspecimentoassaybuffer. Gentlystirtheassaybufferwiththelooptoensureadequatemixing oftheserumintheassaybufferandremovetheloop. Removetheteststripfromthefoilpouchpriortouse. Holdingthestripvertically,inserttheteststripintothetube containingdilutedspecimen. - 111 -

EZLabManual Interprettestresultsat15to30minutes.Ifthetestbandisvery faintat15minutes,pleasereadtheresultsagainat30minutes.

INTERPRETATIONOFTHETEST IgMPositive Thecontrolline(C)andIgMline(M)arevisibleontheteststrip.Thisis PositiveforIgMantibodiestoSalmonellatyphi.THISISINDICATIVEOF ACUTETYPHOIDFEVER. IgGandIgMPositive Thecontrolline,IgM(M)andIgGline(G)arevisibleontheteststrip. ThisispositiveforbothIgMandIgGantibodies.ThisisindicativeofACUTE THYPOIDFEVERinthemiddlestageofsalmonellatyphiinfection. IgGPositive ThecontrollineandIgGline(G)arevisibleontheteststrip.Thisispositive forIgGantibodies.ThisisindicativeofpreviousSalmonellatyphiorrelapseor reinfection NEGATIVE Thecontrollineisonlyvisibleontheteststrip.NoIgGandIgMantibodies weredetected.ProbablyNOTTHYPOID.

- 112 -

EZLabManual SPECIALTEST WIDALTEST Procedure: 1. Deliver0.08,0.04,0.02,0.01and0.005mloftheserumtothesquaresineachrow. 2. Addtoeachrow0.03mlofantigensuspensionsandmixwithapplicatorsticks. 3. Rocktheslideontheilluminatedboxfor3mins.Andreadtheagglutination. 4. Thedegreeofagglutinationisreadinaccordancewiththefollowingscale: Completeagglutination 4+ 75%agglutination 3+ 50%agglutination 2+ 25%agglutination 1+ 5. The serum dilutions on the slide are considered to correspond with the following dilutionsinthetest: 0.08mlserum 1/20 0.04mlserum 1/40 1/80 0.02mlserum 0.01mlserum 1/160 0.005mlserum 1/320 The liter of the serum against the antigen is considered to be shown by that serum givinga2plusagglutination.

- 113 -