SPECIMEN COLLECTION

AND HANDLING
FOR DIAGNOSIS OF INFECTIOUS DISEASES

 Appropriate specimen
collection
handling

= key to diagnosis.

When ?
likelihood of recovering the suspected agent is greatest.
Ex: viruses in the acute phase

BEFORE antimicrobial therapy !

 How much ?
Volume: adequate

Swab
polyester-tipped swab on a plastic shaft is
acceptable for most organisms.
not optimal for:
anaerobes,
mycobacteria
fungi.

 Obtained with minimal contamination

labelled:

name
identification number
source
Required examinations,
Collection time

Sterile container
except stool (clean container = enough)

After collection
placed in a safety bag
transported to the laboratory as soon as possible.

 Delay unavoidable:
refrigerated to prevent overgrowth of the flora

urine,
sputum and other respiratory specimens,
stool,
specimens for Chlamydia trachomatis or viruses

stored at room temperature

CSF and other body fluids,
blood
specimens for Neisseria

RESPIRATORY TRACT
1. Throat swab: pharyngitis, scarlet fever, diphteria,
viral respiratory infections, etc
- When:
- morning,
- before eating and teeth brushing

- How:
-

depressing the tongue with a tongue blade,

swab between the tonsilar pillars and behind the uvula
without touching the lateral walls of the oral cavity,
swabbing back and forth across the posterior pharynx.

- for viruses - viral transport medium,

- for bacteria - modified Stuarts medium.

Secreie faringian

CDT Cluj-Napoca

CDT Cluj-Napoca

RESPIRATORY TRACT

2. Nasopharyngeal specimens:
- aspirates,
- washings,
- swab => for each nostril
Especially for viral respiratory infections
- sinusites,
- rinitis,
- measles,
Bacteria
- Clamydia trachomatis pneumonia in infants,
- diphteria,
- Pertussis
- carriers of Staphylococcus aureus.

RESPIRATORY TRACT
3. Sputum and tracheal aspirates
- bronchitis, pneumonia, tuberculosis
- morning,
- after mouth is rinsed with saline or water.
- 5-10 ml,
- resulting from a deep cough
Tracheal aspirates
- Catheter
- in patients with tracheostomies,
- represents lower respiratory secretions.

A purulent sputum
B- sputum

http://www.microbes-edu.org/glossaire/glossaire.html

EYE
1.Conjunctival specimens: scrapings or swab
- conjunctivitis
2. Corneal scrapings and biopsy
- keratitis
- with a platinum spatula, used for preparation of
smears and culture on media.

DIGESTIVE TRACT
1.

Vomit, food
1.

in food poisoning

2. Feces (stool)
Bacteria, viruses:
- diarrhea, food poisoning, botulism
- Spontaneous defecation
- swab inserted beyond the anal sphincter, rotated, withdrawn
and placed in transport container with a tight lid.
Parasitic protozoa and helminth eggs or larvae.
fresh stool placed in a clean, dry container
-

not collected from the toilet bowl,

not contaminated with urine, antidiarrheal compounds, other
substances

container with Carry Blaire medium

CDT Cluj-Napoca

Leifson medium
Klebsiella spp.
E. coli (down)

Salmonella spp. (up) and

3. Bile
- morning,
- after mouth is rinsed with saline
- for bacteriologic (biliculture) or
parasitologic examinations.

URINARY TRACT
1.Mid-stream clean catch
- first voided morning specimen
2. Catheterisation or suprapubic aspiration
sterile container
transported promptly to the laboratory.
processed within 2 hours after collection.
Indications:
urinary infections (cystitis, etc),
salmonelosis, leptospirosis, tuberculosis.

URINARY TRACT
Quantitative bacterial culture
inoculate appropriate media with a measured
amount of urine,
commonly with a calibrated loop
(0.01/0.001ml)

Nr of mo/ml = nr colonies x 100/1000

on blood agar plate
~ Infection > 100.000 CFU/ml not only !!!

Growth of three or more species =>

contamination.

Quantitative bacterial culture of a urine specimen

calibrated loop

CDT Cluj-Napoca

CDT Cluj-Napoca

E. coli

Mediul Levin

CDT Cluj-Napoca

Mediul CLED

Strips for biochemical analysis - urine

CDT Cluj-Napoca

GENITAL TRACT
1.Vaginal secretions in vulvovaginitis and
bacterial vaginosis
- a wet mount preparation
- 48-72 h after sexual intercourse
2. Urethral specimens
- sexually transmitted disease
- urinary infections
- morning / 2h after voiding

SECRETIE
URETRALA

SECRETIE
VAGINALA

Chlamydia trachomatis

Candidoza urogenitala

BLOOD
Different purposes:
1.Bacteriologic
- bacteriemia,
- sepsis,
- infections of vascular grafts, etc.
2.Serologic: presence and titre of antibodies
3.Virology
4.Parasitology
5.Hematologic
6.Biochemical examinations.

BLOOD
Blood culture
- collected just before a chill ( or after)
-drawn with a needle directly into bottles of culture media
-venal puncture
skin is cleaned with alcohol, iodine.
-from the first bottle with culture media, several second
(blind) cultures on solid media are performed every 2-3
days.
Several semi - to fully automated blood culture systems
available
- eliminate the need for blind subculture
- Short incubation period from 7 to 4/5 days.

http://www.microbes-edu.org/glossaire/glossaire.html

CDT Cluj-Napoca

CDT Cluj-Napoca

BLOOD
Serology

venal puncture,
5-10 ml,
Coagulate => clot removed
serum is used as required.
Serum can be preserved at - 20C.

Parasitologic
malaria, babesiosis, trypanosomiasis and some filariases
Collected from finger

Hematologic
finger or by venopuncture with anticoagulant ( the first method is
preferable).

-wet-mount can be examined

-thin smears => Giemsa stain (or other staining)
-thick smears are useful for detecting all blood parasites.

PURULENT MATERIAL
-aspirated with a needle and syringe from closed
collections or with the loop or swab from the opened
collection after the cleaning of the plague and the
removing of the first quantity.

TISSUES
-obtained surgically for microbiologic and histopathologic
examinations.
-fragments from the corps can be obtained after death.

CEREBROSPINAL FLUID (CSF)

-collected to diagnose meningitis, viral encephalitis
-is obtained by lumbar spinal puncture or by
aspiration from ventricles
Skin antisepsis and temperature ( 37C) are very
important.
-submitted to the laboratory in 3 glass tubes:
-one for cell counts and differential stains,
- one for smears stains with Gram or other stains
and culture,
- one for glucose and protein dosage and other
special tests (serologic).

http://www.nlm.nih.gov/medlineplus/http://www.microbesedu.org/etudiant/etudiants.htmlspanish/ency/images/ency/fullsize/9242.jpg