LABORATORY DIAGNOSIS

Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)

TIMMING OF SPECIMEN COLLECTION

Best time to collect:

ACUTE PHASE of illness

BEFORE antimicrobial therapy

SPECIMEN VOLUME

Should be adequate
If not, prioritize requests

For SWABS:

Polyester-tipped swab on a plastic shaft

What to avoid in collecting swab specimens:

o
Calcium Alginate if collecting for viral culture,
could inactivate HSV
o
Cotton toxic to N. Gonorrhoea
o
Wooden Shaft (Wood) toxic to C. trachomatis

Swabs are not optimal for detection of ANAEROBES,

MYCOBACTERIA or FUNGI

ACTUAL TISSUE or FLUID ASPIRATE is always superior than

swab
SPECIMEN COLLECTION

Site of infection
Sterile container
Include the following (label the container):
o
Name and identification number of Pt
o
Source of specimen (body location)
o
Date and time collected

SPECIMEN TRANSPORT
ALL SPECIMENS MUST BE:

In a biohazard bag

transported ASAP
DELAYS:

REFRIGERATE:
o
Urine
o
Sputum and other respiratory specimen
o
Stool (for detection of C. trachomatis or viruses)

DO NOT REFRIGERATE (keep @ room temp):

o
Blood
o
CSF and Body Fluids
o
Specimens for recovery of N. gonorrhea

UNACCEPTABLE SPECIMENS
The laboratory director must create criteria for rejecting specimens.
Specimens rejected:

Specimen/s in formalin

24 hr sputum

Leaking samples

Specimen inoculated into agar that has dried or outdated

Contaminated by: BARIUM, CHEMICAL DYES, OILY CHEMICALS

Foley catheter tips

Duplicate specimen received w/in 24hr period
Blood catheter tip from patient without concomitant positive
blood culture

REJECTED FOR ANAEROBIC CULTURE:

Gastric washing

Urine other than suprapubic aspirate

Stool (EXCEPT: for recovery of C. difficile)

Oropharyngeal specimen (EXCEPT: deep tissue specimens

obtained during surgery

Sputum

Swabs for ileostomy

Superficial skin specimens

UNIVERSAL PRECAUTION
PPE:

Gloves and lab coat

Masks and goggles
Gowns or aprons

Ideally, all specimens, but at a minimum respiratory specimens and those

submitted for detection of mycobacteria or fungi must be opened in a
BIOLOGICAL SAFETY CABINET
REFERAL TESTING
Packed according to dangerous goods shipping guidelines (Internation
Air Transport Association)

No more than 40 ml

Cultures of bacteria and fungi placed on solid media in tubes

Capped and sealed with waterproof tape

Placed in a secondary container

If several primary container, it must be individually wrapped

or separated to prevent contamination between containers

Secondary packaging/container:
o
must be leakproof, capped and placed in a
corrugated fibreboard or hard plastic
o
must be able to withstand temperature and air
pressures it will be subjected to

Itemized list must be enclosed in between the secondary and

outer packaging

Dry Ice, if needed, must be placed outside second container

All infectious shipping packages must be labelled, containing address
and contents as well as name and address of person responsible for
the shipment
BLOOD
Most important function of microbiology laboratory: detection of blood
borne pathogens
Culture of blood is essential for identifying bacteria responsible for:

Bacteremia

Sepsis

Infections of native and prosthetic valves

Suppurative thrombophlebitis

Mycotic aneurism

Infections of vascular grafts

LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)
Important in Dx of invasive or disseminated infections caused by certain
fungi especially:

Candida spp

Cryptococcus neoformans

Fusarium spp

Histoplasma capsulatum
Parasites are detected using peripheral blood smears
Blood should be collected for culture before antimicrobial therapy when
one or combination of the following are present:

Fever (38 deg or higher)

Hypothermia (36 deg or lower)

Leukocytosis (esp. Left shift)

Granulocytopenia

Hypotension
SPECIMEN COLLECTION OF BLOOD

Patients with endocarditis (optimal number blood cultures for

detection of bacteremia varies due to controversies ):
o
two to three 20mL blood samples drawn over 24hr
period and equally distributed into aerobic and
anaerobic blood culture bottles
o
four blood cultures drawn w/in 24 hours with 30-60
mins interval for the first 2 sets, another one to two
sets drawn over remaining 24hrs if septicaemia
persists (inc yield of potential pathogens by 20%)
o
if antimicrobial therapy initiation is urgent, it should
be collected before therapy begun, from separate
sites within a few mins.

RECOVERY OF MICROORGANISM

Timely collection and accurate identification of organisms depend on:

Appropriate collection

Transport

Processing of specimen
To minimize contamination of normal flora:

Venipuncture site should be prepared with a bactericidal

agent:
o
70% isopropyl or ethyl alcohol
o
1-2% iodine solution
o
Iodophor or chlorhexidine

For maximum antisepsis: dry are for 1-2 minutes before

venipuncture

Host factors that might impede recovery of microorganisms:

antibodies

complement

WBC (phagocytic)
Various mechanisms to counteract host factors:

Diluting blood specimens

o
broth medium
o
1:10 ratio to neutralize serum bacterial activity

Incorporate 0.02-0.05% sodium polyethanol

o
Inhibits coagulation, phagocytosis, complement
activation and inactivates aminoglycosides

Counteract microbial agents:

o
Antibiotic-adsorbent resins
o
Lysis-centrifugation system

APPROPRIATE TIMING FOR DETECTION OF BACTEREMIA AND

FUNGEMIA

BLOOD CULTURE SYSTEMS

Optimal time:

Just before a chill (not predictable)

collected after onset of fever and chills

PROCEDURE: Blood -> bottles of culture media -> inverted several times
to ensure mixing - > transported to laboratory @ room temperature
SPECIMEN VOLUME
Adults 20-30mL blood per culture set
Infants and Children 1-5mL
SPECIMEN DRAWS
Recommended number of blood specimen to collect depend on nature of
bacteremia:

TRANSIENT BACTEREMIA
o
Manipulation of a focus of infection
o
Instrumentation of a contaminated mucosal surface
o
Surgical procedure in a contaminated site
o
Occurs early in course of many systemic and
localized infections

INTERMITTENT BACTEREMIA
o
Associated with undrained abscess

CONTINOUS BACTEREMIA
o
Hallmark of intravascular infection
o
Occurs during first few weeks typhoid fever and
brucellosis

Manual Blood Culture

o
Antibiotic Adsorbent Resins

Biphasic system

Broth medium in a bottle with attached

chamber containing agar media on a
paddle

PROCEDURE: Bottle is tipped -> bloodbroth mixure enters chamber and flow
over agar media -> Colonies from agar
media: used for identification and
susceptibility testing

For AEROBIC and FACULTATIVE

BACTERIA and YEAST
o
Lysis-Centrifugation System

Inhibits coagulation and complement

cascasde

Lyse blood cells

Provide cushion for microorganism

during centrifugation

PROCEDURE: Blood -> tube ->inverted

several times ->transported asap >centrifuged for 30 mins @ 3000g ->
supernatant discarded ->sediment mixed
in vortex mixer -> plated to agar

Smaller tubes for low vol samples

ADVANTAGES:

Excellent for recovery of

Staphylococcus aureus some
LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)

Enterobacteriaceae and
fungi

Best for recovery of H.

capsulatum

Direct availability of colonies

for identification and
susceptibility testing

Carry out quantitative cultures

Flexible, special media can be

inoculated to recover
organisms with specific growth
requirements (eg. Legionella
and mycobacteria)

DISADVANTAGE:

Labor intensive

Less likely to recover

Streptococcus pneumoniae,
Haemophilus influenzae or
anaerobes

Risk of contamination is
incresed
Automated Blood Culture
o
Based on calorimetric detection of CO2 produced
during microbial growth

PROCEDURE: inoculated bottles

continuously rocked -> if bacteria is
present ->production of CO2 -> released
to broth -> pH dec -> sensor change color
-> green to yellow

Medium available:

Routine: 5-10mL blood

PediBacT: </= 4mL blood

Fastidious antibiotic neutralization

enhances detection of fungi and bacteria
from Pt under antimicrobials
o
Based on fluorescent technology

CO2 sensor generates hydrogen ions ->

dec pH -> inc fluorescence -> change
signal -> computer generates growth
curves -> data analyzed via growth
algorithms

Medium available:

Aerobic and anaerobic low vol:

5-7mL blood

High vol: 8-10mL blood

Peds Plus: 0.5-5mL blood

Myco F Lytic: recovery of fungi

and mycobacteria
o
Based on measuring gas consumption

PROCEDURE: inoculated vial fitted with

disposable connector that contains
recessed needle -> penetrates bottle
stopper and connects bottle headspace to
sensor probe -> sensor monitors
consumption and or production of all
gases (CO2, N2, H2) -> creates data points
in computer

Medium available:

Aerobic and anaerobic media:

80mL broth + 0.1-10mL blood

EZ Draw (direct draw): 40mL

broth + 0.1-5mL blood

DETECTION AND NOTIFICATION OF POSITIVE CULTURES

Positive blood cultures with commonly isolated aerobic

organisms are detected within 1236 hours of incubation.
o
The initial report is a Gram stain report only.
o
Identification and susceptibility report w/in 2448
hours after the Gram stain report.

Cultures containing anaerobes;

o
not detected for 4872 hours
o
Identification is not available for 34 days after that.

Fastidious organisms, such as those found in the HACEK group

may not be detected until 35 days.

Haemophilus

Actinobacillus

Cardiobacterium

Eikenella,

Kingella)

Detection of Viruses

Blood specimens are most commonly collected to monitor

response of infection to antiviral therapy by using
quantitative polymerase chain reaction (PCR) to measure
viral load of:
o
human immunodeficiency virus (HIV),
o
hepatitis C virus (HCV)
o
hepatitis B virus (HBV)
o
cytomegalovirus (CMV)

For HIV and HCV, blood specimens also may be collected for
genotyping

PCR (qualitative or quantitative PCR) generally is used to

confirm an initial positive HCV antibody result.

In addition to assessing response to antiviral therapy,

measuring viral load in a blood specimen is useful for
monitoring for development of disease and for diagnosis of
disease in specific situations.

determining the level of CMV deoxyribonucleic acid (DNA) in

blood is used to predict those at high risk for development of
CMV disease and direct the initiation of pre-emptive therapy
in;
o
immunocompromised patients
o
transplant recipients
o
acquired immunodeficiency syndrome (AIDS)
patients,

Monitoring the level of Epstein-Barr virus (EBV) DNA in serum

or plasma by quantitative PCR is indicated in transplant
recipients at high risk for EBV-associated lymphoproliferative
disease.

Quantitative PCR from whole blood or peripheral blood for

diagnosis of HHV-6 or HHV-7 (both causative of roseola
infantum or exanthema subitum) on transplant recipients and
is the test of choice for diagnosing disease caused by
parvovirus B19 in immunosuppressed patients or in the fetus.

LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)
Table 63-2 -- Infectious Meningitis Syndromes

Detection of Parasites

Blood specimensare useful for diagnosis of:

o
malaria,
o
babesiosis,
o
trypanosomiasis,
o
some filariasis.
Specimens should be collected in tubes with anticoagulant and
transported promptly to the laboratory
If smears must be sent to a reference laboratory, they should
be fixed in absolute alcohol soon after they are made.
The simplest technique for detecting parasites in a sample of
blood is the direct mount, prepared by placing one drop of
blood on a glass slide, covering it with a cover glass, and
examining it immediately.
Direct mounts are excellent for diagnosis of trypanosomiasis or
filariasis because the trypomastigotes and the microfilariae
easily can be seen moving, often with low or medium power.

Syndrome Onset/Duration

Probable Pathogens

Acute

< 24 hours

Pyogenic bacteria

Subacute

17 days

Enteroviruses, pyogenic bacteria

Chronic

Persisting at least 4
weeks

Mycobacterium tuberculosis
Treponemapallidum
Brucella sp.
Leptospirainterrogans
Borreliaburgdorferi
Cryptococcus neoformans
Coccidioidesimmitis
Histoplasmacapsulatum
Candida sp.

The definitive diagnosis is made by staining smears

The thin smear, made as for hematologic work and stained in a

similar manner, is the standard preparation for determining
the species of
o
Plasmodium,Babesia,
o
Trypanosoma,
o
microfilaria
Thin smears for parasitologicwork are fixed and then
preferably stained manually with Giemsastain, but automated
hematologic staining is adequate.
Smears are first scanned at low power to detect microfilariae,
which are large objects (between 100 and 200 m) and easily
seen, usually at the lateral edges of the smear.
After they are located, microfilariae should be studied under
oil immersion for identification.
Following scanning with low power, thesmear is examined
with a high dry objective, searching for trypanosomes;
And finally under oil immersion, to find and identify
Plasmodium, Babesia, and Trypanosoma.
Thick smears are useful for detecting all the parasites
mentioned earlier, and are part of the minimum laboratory
work-up for their diagnosis.
1. A drop of blood is placed on a clean glass slide
2.
with the corner of another slide, gently spread to
cover 1 cm square.
3.
The preparation is allowed to dry
4. Without fixation is stained with Giemsa stain,
allowing for its dehemoglobinization.

CEREBROSPINAL FLUID

CSF is collected to diagnose meningitis and, less frequently,

viral encephalitis.

Infectious meningistis

A medical emergency requiring early therapy to prevent death

or serious neurologic sequelae
Divided into acute, subacute& chronic clinical syndromes
based on duration of symptoms.

The enteroviruses are the agents most commonly responsible

for meningitis, and they should be considered first in the
differential diagnosis of meningitis in a child or adolescent
during the late summer and early fall.
The pyogenic bacteria responsible for meningitis vary with
the age of the affected individual

Table 63-3 -- Common Bacterial Causes of Acute Meningitis by Age

Age

Organisms

Neonates3 months Group B streptococcus

Escherichia coli
Listeria monocytogenes[*]
Streptococcus pneumonia
4 months6 years[] Streptococcus pneumonia
645 years

Neisseria meningitides

> 45 years

Streptococcus pneumonia
Listeria monocytogenes
Group B streptococcus

May cause meningitis in immunocompromised individuals in all age

groups.

SAMPLE COLLECTION AND TRANSPORT

CSF is usually obtained by lumbar spinal puncture

sometimes it is aspirated from the ventricles or collected from
a shunt
As when collecting blood for culture, careful skin antisepsis is
essential for collection of CSF, which typically is submitted to
the laboratory in three tubes

Suggestions for tests performed on fluid in each tube are:

o
o
o

Tube1 - protein and glucose

Tube2-preparation of smears to stain with the
Gram's stain or other stains and for culture
Tube3-cell counts and differential stains

LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)
o

Tube4- only if indicated, special tests such as the

cryptococcal antigen, serologic test for syphilis,
other serologic studies, and cytology.

Table 63-4 -- Normal Cerebrospinal Fluid Parameters and Changes

in Infectious Meningitis
Condition

WBCs (cells/L)[*]

Protein
(mg/dL)

Glucose
(mg/dL)

Normal

5 (lymphocytes)

1445

45100
(2/3 serum)

500200 000 (PMNs)

Meningitis
Acute/subacute
bacterial

Chronic
bacterial 2002000
tuberculous, fungal
(lymphocytes)
Enteroviral

2002000
(PMNs
early;
lymphocytes
later)

Mycobacteria

Detected only if the findings in the CSF shows

pleocytosis(increased lymphocyte with decreased glucose in
CSF), or decreased glucose, or elevated protein values

For optimal recovery, culture of at least 5mL is recommended

Fluid is centrifuged at 3000-3600g for 30mins

Supernatant is decanted

Sediment is mixed on vortex mixer (to prepare smears and to

inoculate appropriate media)

Mycobacterium tuberculosis: nucleic acid amplification for

direct detection
Processing CSF for detection of fungi

Similar to that of bacteria

Organisms are concentrated by filtration or centrifugation

Gram stain

Media: Brain-heart infusion or SABHI agar without antibiotics

Normal

= increased; = decreased; PMNs = polymorphonuclear leukocytes;

WBC = white blood cells.

ADDITIONAL DIAGNOSTIC TESTS

Latex agglutination test

CSF should be transported promptly to the laboratory and

processed as rapidly as possible
If a brief delay in processing is unavoidable, the specimen
should be held at room temperature unless viral culture is
requested, in which case a portion (preferably 1 mL but no less
than 0.5 mL) may be refrigerated for a short time
Specimen processing differs for bacteria, fungi, viruses, and
parasites and is discussed separately for each group of
organisms.

SAMPLE PROCESSING FOR BACTERIAL AND FUNGAL CULTURE

Processing of CSF (routine bacterial culture)

Includes concentration if 1mL or more of specimen is received

Preparation of smear by cytocentrifugation (Gram stain)

Culture
Concentrate the fluid by centrifugation of 1500g for 15mins

Supernatant decanted into sterile tube

Leave 0.5mL of sediment and fluid mixed on vortex mixer, or

forceful aspiration

May be performed with the use of the supernatant of a

centrifuged specimen or the original fluid
For detection of antigens of ;
o
Steptococcus agalactiae,
o
S. pneumoniae
some stereotypes of ;
o
Neisseria menigitidis,
o
Escheria coli
o
H. Influenza type b.
Most useful in diagnosing partially treated meningitis and
confirming a positive Gram-stained smear
Disadvantage: Compared to gram stain, its sensitivity is not
significantly greater and more expensive

2 Tests for Diagnosis of Menigitis caused by C. Neoformans:

1.

Chronic bacterial meningitis

Requires special requests

India Ink preparation

Allow visualization of encapsulated yeast cells (Fig.

1)

Performed by mixing 1 drop of CSF sediment with 1

drop of India ink

Sensitivity is low, except in HIV-infected persons

Brucellosis

Routine bacterial culture

Media are incubated for 2-3 weeks

Leptospirosis

Leptospira interrogans may be cultured during the first few

weeks of illness
Neurosyphillis

Pleocytosis

Elevated protein concentration

Positive VDRL test (only useful method for detecting

antibodies to Treponema pallidum in the CSF)
B. burgdorferi (Lyme Disease)

Involvement of CNS

Detection of IgM and IgG in antibodies in CSF and serum

Fig. 1. India ink preparation of CSF fluid shows encapsulated yeast forms
of Cryptococcus neoformans.
2.

Latex agglutination and ELISA

LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)

Specific for capsular antigen

Highly specific and have sensitivities of more than
90%,
Recommended for diagnosis
Supernatant of a centrifuged specimen or unspun
CSF can be used
False-positive latex agglutination results occur due
to the presence of Trichosporon asabii or to the
introduction of trace amounts of condensation form
agar into the test fluid
Should be performed before culture or on separate
sample to avoid problem

To diagnose peritonitis associated with chronic ambulatory

peritoneal dialysis, collection of at least 50 mL of fluid may
improve recovery of the responsible pathogen.
To transport the fluid, it is aspirated into a sterile container
and delivered promptly to the laboratory.
Enteroviruses, primarily Coxsackie viruses A and B, are among
the most common causes of infectious pericarditis
Collection of throat washings and stool (which are more likely
to yield the virus), in addition to pericardial fluid, is strongly
recommended for viruse isolation from persons with
suspected enteroviral pericarditis.

Sample Processing for Bacterial Culture

Sample Processing for Diagnosis of Viral and Parasite Infections

Nucleic acid amplification tests (NAAT) are used most often

for diagnosis of viral infections of the CNS.
Conventional cell culture (primarily for detection of
enteroviruses although PCR is preferred)
Serologic tests for viruses that cause encephalitis (western
equine, Venezuelan equine, St Louis, Japanese, and La Crosse
and West Nile.
CSF is occasionally sent to the laboratory for diagnosis of
o
African trypanomiasis;

Trypanosoma gambiense

Trypanosomarhodesiense
o
infection with free-living amoebae

Naegleriafowleri

species of Acanthamoeba
Specimen received in the laboratory should be processed
immediately
Wet preparations are prepared directly from the specimen and
from the sediment, by first shaking the tube gently and the
centrifuging the specimen at 250g for 10 min.
Preparations are examined under the microscope with the
condenser in a low position to allow visualization of
trophozoites or by phase contrast microscopy (preferably).
Cultures of free-living amoebae from CSF are:
Done on non- nutrient agar plates covered with a
suspension of E.coli or Enterobacteraerogenes.
fluid is centrifuged at 250 g for 10 min
Supernatant is removed with a sterile pipette
Sediment is mixed with 0.5mL of saline solution and
poured at the center of the plate
Culture is incubated at 37C and examined for amoebas
daily for 10 days using a microscope under a 10x
objective

PARASITOLOGICAL EXAMINATION

OTHER BODY FLUIDS

Fluid is collected by aspirating using a needle and syringe from

the
Pericardial cavity
Thoracic cavity
Peritoneal cavity
Joint spaces
A volume of 1-5mL is adequate for isolating most bacteria, but
10-15 mL is optimal for recovery of mycobacteria and
fungi

Processing fluid from body cavities for detection of

bacteria involves preparing a smear for Gram stain and
inoculating appropriate media for culture
In the laboratory, the fluid is centrifuged at 1500-2500 g
for 20-30 min.
The supernatant is removed, leaving about 0.5mL fluid in
addition to the sediment, which is mixed thoroughly and
then used to prepare smears and inoculate media.
Fluid specimens submitted for detection of mycobacteria
are processed.
Fluid for fungal culture should be concentrated by
centrifugation as described for bacteria.
The supernatant is removed, leaving 1.5-2.0mL, in which
the sediment is thoroughly mixed.
A smear of the sediment is prepared for staining with
Gram stain or Calcofluor white.
0.5-1.0 mL of sediment is inoculated to primary fungal
planting media (as for CSF), but lesser volumes are
acceptable.

Body fluids are rarely collected for detection of parasites

Entamoeba histolytica may be found in the pericardial,
pleural, or peritoneal cavity
o
result of rupture of an abscess of the liver into the
peritoneal, pleural, pericardial cavity
o
the lungs: into the pleural or pericardial cavity
o
perforation of amebic ulcers into the peritoneal
cavity
Hydatid cysts (Echinococcus spp.) are infrequently
diagnosed by examination of body cavity fluid, also due to
rupture of a cyst into a viscus contiguous to the cavity in
question.
o
The fluid collected is usually clear and contains
hydatid sand but rarely is turbid because of
superimposed bacterial infection.
Filarial infection examination of wet preparations of a body
cavity fluid may demonstrate the microfilariae
Strongyloides hyperinfection larvae may be detected in body
cavity fluids
TISSUE

Tissue specimens obtained surgically are procured at great

expense and at considerable risk to the patient; therefore, it
behooves the surgeon to obtain an amount of material that
LabDiag Trans Group 2017

LABORATORY DIAGNOSIS
Chapter 63/Lecture 4
Specimen Collection and Handling of Infectious Diseases part 1
(Blood, Body Fluids, Tissue)

is adequate for both histopathologic and microbiological

examination
The histopathology of the lesion serves not only to
differentiate between infection and malignancy but also to
distinguish between a suppurative and a granulomatous
process.
In some cases, special stains are helpful in establishing the
etiology of the process.
In chronic lesions the differential diagnosis includes disease
due to actinomycetes, brucellas, mycobacteria, and fungi, any
one of which may be present only in small numbers

SAMPLECOLLECTION AND PROCESSING

Tissue obtained surgically for culture should be placed into a

sterile, wide-mouthed, screw-capped container
As a general rule, tissue should be bisected aseptically by the
surgeon in the operating room and material representative of
the pathologic process should be submitted for both
histopathologic and microbiological examination.
Tissue received in the laboratory is finely minced with sterile
scissors or scalpels, added to a small volume of broth, and then
rendered homogeneous either in a tissue grinder, mortar and
pestle, or stomacher to provide a 20% suspension.
This suspension is used to inoculate all of the necessary culture
media and is then stored under refrigeration for at least 2
weeks before being discarded.

4.

5.

Which of the following is true about the most common

etiologic agent of meningitis?
a.
Mostly due to gram-negative bacteria
b.
pyogenic bacteria responsible for meningitis vary
with the age of the affected individual
c.
In the laboratory, the fluid is centrifuged at 15002500 g for 20-30 min.
d. Nucleic acid amplification tests (NAAT) are used
most often for diagnosis of viral infections of the
CNS.
Which is not true about specimen collection and handling of
infectious disease?
a.
Specimen collection is best during fever and chills
b. All specimens must be in a biohazard bag during
transportation.
c.
Venipuncture site should be prepared with a
bactericidal agent
d. All specimen that will not be brought to the
laboratory immediately should be put into an SPS
containing transport media

ANSWERS: DABDD

REFERENCES:
1.
McPherson et al. 2012. Henrys Clinical Diagnosis and
Management by Laboratory Method. 22th ed. Pp. 1239-1244

MCQs
1.

2.

3.

Ms. X is complaining of a lower abdominal pain that become

more pronounce upon urination, vaginal discharge shows
gram-negative kidney bean diplococci, culture was grown on
a Thayer-Martin agar. Which of the following is the LEAST
likely to be used upon specimen collection and handling?
a.
Endocervial specimen can be used
b. Use of CO2 generating tablet on the agar container
c.
Use of first voided urine samples in men
d. Regular swab can be used
e.
Brush can be used for the cervical os
Which of the following specimen collection will mostly be
unacceptable?
a.
induced sputum with >10 epithelial cell
b. Suprapubic aspirate for urine culture
c.
Bronchoalveolar lavage (BAL) lower respiratory
specimen
d. Throat swab for URT specimen
Which of the following statement is TRUE?
a.
Iodine is used on wet mount of body fluids for
detection of pathogenic amoeba
b. NSS can be used to observe moving trophozoite
c.
Dacron swabs are toxic to N. gonorrhoea
d. Supernatant of centrifuged CSF is used for gram
stain and culture
LabDiag Trans Group 2017