lOMoARcPSD|3566749

Finals-LAB- Complete - finals

Medical Technology (Far Eastern University)

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PROCEDURES FOR IDENTIFICATION OF STREP. SPP., ENTERO SPP., GRAM (-) COCCI
A. Cultural Characteristics
1. Color of colonies
2. Size of colonies
3. Type of hemolysis
B. Microscopic Examination
1. Gram Stain
 Cocci; in chains [Streptococci]
 Pairs [Diplococci]
 Kidney-bean shape [Neisseria]
C. Biochemical Characteristics
1. Catalase test
 (–) = Streptococci
 (+) = Neisseria or Moraxella or Gram (-) cocci
2. Taxo A/Bacitracin
 Specimen must be β-hemolytic
 S =Group A Streptococci
 R = other Streptococci
3. Taxo P/Optochin
 Must be alpha hemolytic and Catalase (-)
 Active Chemical Component: Ethylhydrochaprin hydrochloride
 S = Pneumococci >14 mm
 R = Viridans streptococci
4. Taxo N/Oxidase
 For identifying Neisseria spp.
 Tetramethyl-paraphenylene dihydrochloride
 (+) = blue/black-purple
5. CAMP test (Christie Atkins Munch Petersen Test)
 Must be β-hemolytic
 Uses a known organism (S. aureus) and an unknown organism (S. agalactiae or Group B Streptococci)
 Unknown organism is β-hemolytic, Catalase (-), Pinpoint and Bacitracin Resistant)
6. Bile Esculin Test
 Differentiates Group D Streptococci or β-hemolytic
 Enzyme: Esculinase
 (+) = blackening of the medium
7. Salt Tolerance Test
 Uses 6.5% NaCl
 This test identifies what type of Enterococci it is
 (+) = turbidity [Enterococci]
 (–) = clear [Non-enterococci]

URINE AND STOOL CULTURE FOR THE I.D. OF GRAM (-) BACILLI
Urine Culture:
 BAP
 MAC
Stool Culture:
 BAP  XLD
 EMB  SSA
Procedures for ID of Gram (-) Bacilli
A. Cultural Characteristics
B. Biochemical tests
a. CHO Utilization test ~ TSI d. H2S production ~ SIM, TSI, LIA
b. Motility Test ~ SIM e. Citrate Utilization Test ~ SCA
c. Indole production ~ SIM f. Lysine deamination test ~ LIA

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g. Lysine decarboxylation test ~ LIA i. Vogues Proskauer Test ~ MRVP

h. Methyl Red Test ~ MRVP

WATER BACTERIOLOGY
 Used to test for Coliform bacteria

2 methods for testing:

1. Pour plate method (Serial Dilution)
a. Dilute sample in sterile distilled water
b. Prepare 3 tubes:
 1:100
 1:1000
 1:10000
c. Add 1mL distilled water in all 3 test tubes
Add 1 mL specimen to test tube 1:100
Add 1 mL specimen to test tube 1:1000
Add 1 mL specimen to test tube 1:10000
* get from the previous test tube
d. Discard last 1 mL from test tube 1:10000
e. Put the liquid from test tubes onto petri dish, with label.
f. Mix with Nutrient Agar
g. Incubate and count colonies.
# of colonies x 100/1000/10000
2. Most Probable Number Method
 Durham tube for gas formation of bacteria that utilize lactose in the medium
 Presumptive test
a. Stir water
b. Pipette 10 mL for each of the 5 test tubes
c. Incubate for 24 hours @ 35ºC
d. Check the presence of gas formation accompanied by turbidity
e. Count tubes with gas formation
 Completed test
a. Gram stain
b. Biochemical tests

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