Clinical AND ADVANCE pathology

TOPIC: DIAGNOSIS FOR INFECTIONS

LABORATORY DIAGNOSIS FOR STI AND HIV/AIDS 5.02

1. WHAT ARE
THE OPTIMUM SAMPLES AND APPLICABLE DIAGNOSTIC TEST FOR THE FOLLOWING DISEASE? WRITE THE SIGNIIFICANCE AND LIMITATIONS OF
EACH DIAGNOSTIC TEST. TABULATE YOUR ANSWER.
Infection specimen Diagnostic test Advantage disadvantage
Gonorrhoea Genital, rectal, Microscopy Sensitive and specific in
oropharygeal, symptomatic with male urethral
conjunctival swab, discharge, low cost
genital discharge
Culture Sensitive and highly specific in Strict sample collection,
optimized circumstances, transportation and storage
inexpensive and allows are crucial in maintaining
antimicrobial susceptibility viability
testing, the only method that
allows anti-microbial
susceptibilty

NAATs Superior sensitivity compared to

culture, especially for pharyngeal
and rectal samples
Syphilis blood sample drawn Dark-field microscopy The dark-field technique can be
from a vein in your arm; practised
sometimes, a scraping and the microscope optics optimized
by using
from a chancre in the
specimens obtained along the gum
affected area; less margin, inside
commonly, cerebrospinal the mouth. Gingival epithelial cells
fluid taken via a spinal and oral bacteria,
tap, depending on your including spiral organisms can be
clinical presentation seen as bright,
white bodies against a black
background
Suboptimal In the diagnostic
of urogenital samples,
expensive
specialized microscope
fitted with a dark-field
condenser is required, together
with adherence to strict
technical conditions in order
to produce reliable results, the
test should be limited to
specialist laboratories

Direct fluorescent Unfortunately, the specific

antibody (DFA) test Both fluorescein conjugate is
the specificity and sensitivity of the not commercially available in
DFA test are superior many countries
to that of dark-field microscopy, .
especially if monoclonal
antibody is used to make the
fluorescein conjugate,
because the DFA technique
eliminates confusion
with other spiral organisms and a
small number of
fluorescein-stained treponemes are
more easily detected
in the stained smear than in the
unstained preparation
(5).

however, the tests cannot be

The polymerase chain reaction (PCR) recommended for the routine
Nucleic acid test can detect detection of T. pallidum in
amplification tests for DNA equivalents of <10 organisms in blood—even in primary and
T. pallidum a specimen, by secondary disease owing
amplifying specific gene segments to the presence of PCR inhibitors.
from T. pallidum
genomic DNA.

Genetic

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 Patho 2 STI AND HIV/AIDS

Analyzer to detect the amplicons has

improved the
analytical sensitivity of a polA PCR to
approximately one
organism per PCR reaction.
applied to the diagnosis of

congenital syphilis
and also neurosyphilis
antigen is not stabilized and a
suspension must be freshly
prepared on the day of use.
VDRL remains the test of choice
Venereal Disease for the detection of reagin in
Research Laboratory cerebrospinal fluid (CSF), patients
(VDRL) with suspected
Test neurosyphilis.

use of a stabilized antigen, the use of

cards instead
Rapid plasma reagin of slides, and the addition of charcoal
(RPR) test particles to the
antigen as an indicator of but it has recently
flocculation. been superseded by tests that
are more sensitive,
technically less demanding, and
“gold standard” treponemal far less subjective.

good quality
Fluorescent Treponemal
fluorescence microscope,
Antibody
experience in reading, quality
Absorption (FTA-Abs) test
reagents, and an appropriate
dilution of the conjugate
are all critical to the reliability of
the test

Unfortunately, both false-

positive and false-negative
FTA-Abs results are common due
to laboratory error and
subjective reading of the test

Treponemal western blot used as a

(WB) assays confirmatory test for treponemal
antibodies in the serum
of patients with syphilis. I
Chlamydia Urine, swabs in the Tissue culture Specificity 100%
vagina, penis mouth and Sensitivity ranges from 60-90%
anus
Non amplified tests

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Enzyme immunoassay Sensitivity and specificity of 85% Could produce false positive
(Chlamydiazyme) and 97% results
Useful for high volume screening

Nucleic acid Sensi: 75-100%, speci: >95%

Hybridization (NA Detects chlamydial ribosomal
probe) – Gen-Probe RNA
Pace 2 Able to detect gonorrhoea and
chlamydia from one swab
Need for large amounts of
sample DNA

DNA amplification test

PCR, ligase chain Sensi: 95% (PCR), 85% (LCR)

reaction Speci: 100%
LCR ability to dectec chlamydia in
first void urine
Herpes Skin mucosal lesions Tzanck smear, Inexpensive, Sensi and Specifi: Fresh lesion
Biopsies papaniclaou, LOW Suboptimal sensi and specifi
Conjucticval/corneal romanovsky stain
smears

Smear tissue section, Direct Inexpensive, Rapid (4 hours Fresh vesicle, suboptimal
smear from base of immunofluorescence possible) sensitivity, time consuming,
vesicle Typing possible, high sensitivity labour intensive, not
standardized

Swab of exudate from Immune peroxidase High specificity >90%, Reagent Fresh vesicle, suboptimal
the base of vesicle staining cost, rapid, does not require sensitivity
integrity of the specimen, typing
possible

Swab, skin lesions, HSV isolation Gold standard, simplicity of Less sensitive than PCR,
vesicular fluid and susceptible cells (virus sampling, virus typing, resistance storage and transport is
exudate, mucosal culture) phenotype testing crucial before testing, labor
sample, biopsies, intensive, needs specialized
conjunctival corneal, laboratories, 2-7 days result
neonates
HIV Serum, plasma, whole Enzyme Sensitive and specific in detecting Do not detect very early Ab
blood Immunoassays HIV Ab responses in the form of IgM

HIV Rapid Tests Take up to 2-4 hours, the devices
are optimized to accelerate
antigen antibody interaction
Used in setting with low volumes
of specimens to provide more
cost effective care
Requires the use of a high
concentration of antigen and
detection of antigen
antibody complexes with
sensitive colour reagents

4th generation is able to detect Follow up by testing at 4

p24 Ag, w/c can reduce the weeks or later to ensure
window period and identify serocnversion
people who are in acute phase of
infection

Confirmatory assays

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Expensive and require

interpretation to establish a
Western blot Confirmatory test for HIV pos diagnosis based on a
Line immunoassays screening, combination of antigen and
specific antibodies
Detection of p24 Ag, These strips are probed with
RNA and proviral DNA serum specimens. The bound,
HIV-specific antibodies are
detected by secondary
antibodies that are conjugated
with an enzyme followed
by a substrate that yields a
coloured product.

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