Interpretation of Serology Tests in Selected Infections

Interpretation Of Serology Tests In Selected Infections
Prof. Abdulkarim AlAska

Interpretation Of Serology Tests In Selected Infections 1
Antibodies (ABs)
IgM Acute infection IgM cannot cross the placenta
IgG previous infection or exposure IgG can cross the placenta. IgG titre acute phase + Convalescence phase (usually 2 weeks) is needed to confirm diagnosis Sero conversion >2 to 3 weeks but may be delayed.
2
False Negative Serologic Test

1. 2.
3.
4.
Immune system not intact Delay in Antibody response (Lyme disease - Legionnaires Disease) Competition for Antigen binding site of antibody) IgM binds to the Antigen IgG site IgG binds to the Antigen IgM site Prozone Phenomena
3
False Positive
1. 2.
3.
Cross reacting antibody Cross reactivation of latent organism (Influenza Virus A infection activate CMV IgM production Presence of Rheumatoid factors RF = IgM RF + IgG = Complexed = False positive organismspecific IgM Antibody
4
Serological Diagnosis Of Syphilis

I. II. III.
Specific Anti- treponemal Antibody Anti treponemal Antibody Reagin Antibody (BFP)

Test for specific Anti - treponemal Antibody 1. Absorbed fluorescent treponemal antibody (FTA - ABs) 2. Treponema Pallidum Immobilization Test (TPI)
A. B.
C. D.
Most sensitive Utilize living Treponema maintained by passage in rabbits testes. Expensive Potentially hazardous.
6

3.
Treponema pallidum haemagglutination (TPHA) test.

A.
B.
C. D. E.
Sheep, chicken or turkey RBCs. Sensitized by attaching killed Treponema pallidum. Agglutinate by presence ofantibody Less sensitive than FTA Abs Less reliable in the diagnosis of primary syphilis. Sometimes false positive
Serological Tests Of Syphilis

Anti treponemal Antibody Anti-treponemal ABs group detected by Reiter Protein Complement Fixation Test (RPCFT)
A. B.
C.
Appears later than specific ABs Some syphilis patient do not produce the form of ABs Used is limited.
8
Tests For Reagin Antibody
A.
Biological False Positive Antibody (BFP) Reagin Antibody: associated with other diseases (BFP) Acute:
Pneumonia Vaccination with live attenuated viruses. Malaria Pregnancy Leprosy the only infection
B.
Chronic:
Reagin titre falls rapidly with treatment

9
Tests For Reagin Antibody
A large numbers of tests for Reagin:

VDRL
(Venereal Diseases Reference Laboratory). RPR (Rapid Plasma Reagin) ART (Automated Reagin Test)
Good sensitive screening Titre falls rapidly with treatment
Reagin titre falls with treatment.
10
Active Treponema Pallidum Infection

1. 2.

Positive Specific Tests e.g. TPHA Positive ( 1/ 32) of non-specific test (VDRL)
TPI-T (Treponema Pallidum Immobilization Test) FTA T (Fluorescent Treponema Test) Sometimes needed for confirmation.
11
Mycoplasma Pneumonia
Gradual onset, headache, fever, malaria, most typically dry cough. Non respiratory:
Meningitis Encephalitis Pancreatitis Steven Johnsons Syndrome Sensori neural hearing loss
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Mycoplasma Pneumonia
ELISA:
More sensitive and specific Detect IgM, IgA IgM rise early after onset Peak is 1 4 weeks Decline in 2 4 months IgM rise in young patient (Good for diagnosis in young patient)
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IgG and IgA
IgG:
Rise slower. Remain elevated for long time. Rising titre diagnosis (2 samples at least 2 weeks apart indicate current infection)
IgA:
High level in elderly May be more useful than IgM in old patient.
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Legionella
Sensitivity Specificity Technique Description Processing Time Disadvantages
Culture
Growing of bacterium from clinical sample, such as sputum, on specialized culture media Screening of urine sample for the presence of specific legionella antigen (cell markers)
80 %
100 %
3 5 days
Requires that laboratory technicians have specialized training and expertise. Will only diagnose infections with L. pneumophila serogroup 1
Urinary Antigen Test
80 %
95 %
Within hours
15
Cont. Clinical Diagnosis

Sensitivity
Specificity
Technique
Description
Processing Time
Disadvantages
Direct fluorescent antibody (DFA) stain of sputum or other sample from lung
Visual screening of sputum or other sample from lung for legionella bacteria; screening is done under a UV microscope, using fluorescently tagged antibodies to light up bacteria
33 70 %
95 100 %
Within hours
Easy to miss bacterium on microscope slide; results difficult to interpret; requires that laboratory technicians have specialized training and expertise
Screening of blood sample for antibodies to legionella; generally 40 Antibody requires comparison of 60% testing results from two samples, (serology) one collected during acute illness and the other 2-8 weeks later Interpretation Of Serology Tests In Selected Infections
95 100 %
2-8 weeks
Sensitivity is low; for optimal results, requires collection of second blood sample. 16
Legionella Urinary Antigen Tests

I. II.
(Simple, rapid) (70 80 % sensitive) ( 80 100 % specific)

ELISA, needs machine Paper Chromatography
(No instrument ,30 minutes)
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Serological Tests Used In The Diagnosis Of Human Brucellosis

1. 2. 3.
4.
5.
Serum Agglutination Test (SAT) 2-Mercaptoethanol Coombs Test Microplate Agglutination Test ELISA Test OTHERS:

Complement Fixation Test Rose Bengal Test Gel Precipitation

18
1. SAT = STAT (Standard Tube Agglutination Test)

IgM, IgG, IgA Prozone End Point Agglutination (1 / 80 ?, 1 / 160 ?, 1 / 320) Persisting Antibody up to 24 months.
19
2. ME (Mercaptoethanol)
Remove IgM Persisting IgG + IgA is diagnostic for persistent relapsing of Brucellosis. Decrease IgG is prognostic of successful outcome
20
Brucellosis
A.
COOMBS TEST
Detect non-agglutinating antibody (Adding AHG)
B.
MICROPLATE AGGLUTINATION (Antigen + NaCl + AHG)

Highly sensitive End Point agglutination not defined
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S. Typhi
H Antibody Non specific O Antibody Most lab. 1 / 80
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Causes of Negative Widal Agglutination Test

The carrier state An inadequate inoculum of bacterial antigen in the host to induce antibody production Technical difficulty or errors in the performance of the test. Previous antibiotic treatment Variability in the preparation of commercial antigens.
Causes Of False-positive Widal Agglutination Tests

Previous immunization with Salmonella antigen. Cross-reaction with non typhoidal Salmonella. Variability and poorly standardized commercial antigen preparation. Infection with malaria other enterobacteriaceae charring the same s-LPS .
H Pylori
TEST EFFECT OF H2 DRUGS ANTI -BIOTIC REMARKS
Antibody IgM, IgG, IgA
NO
NO
Population study Past or current infection

Active Infection Active Infection
25
Urea Breath Test Fecal Antigen
YES +
YES YES
Toxoplasmosis IHA Test
APPLICATION: To detect Toxoplasma antibodies by indirect haemagglutination test. INTERPRETATION OF RESULTS: Results will be reported as:
A. B.
C.
Positive Doubtful Negative
Doubtful results should be retested within 2 weeks.

26
Comments
There is evidence that very early infectious and those of infants under 1 year of age may not be detected by TOXO IHA Test.
1.
2. 3.
Titres of 1:64 to 1:128 are usually indicative of past exposure, and can be the result of an asymptomatic infection. Titres of 1:256 are usually indicative of a recent infection, not necessarily of a still active infection. Titres of 1:512 and greater indicative of Toxoplasmosis.
In ocular Toxoplasmosis, titres of antibodies may be very low.

27
Toxoplasma IgM Elisa
APPLICATION: For measurement of the IgM antibodies to toxoplasma gondii in human serum and plasma to aid in the diagnosis of primary infection. INTERPRETATION OF RESULTS:
A. B. C.
Negative : Equivocal : Positive :
< 0.500 (arbitrary units) 0.500 - 0.599 0.600.

28
Comments
Transplacental transmission of the parasite resulting in Congenital Toxoplasmosis can occur during acute acquired maternal infection. The risk of fetal infection is a function of the time at which acute maternal infection occurs during gestation. Maternal infections acquired before conceptions present very little, if any, risk to the fetus
29
Comments (2)
Prospective studies of pregnancies have shown that prenatal diagnosis if infection followed by prenatal therapy reduces the frequency and Severity of Congenital Toxoplasmosis.
30
Comments (3)
Since persisting IgM levels may be detected long after the onset of acquired infection, the use of a single serological test result must be used with caution in those cases when it is critical to establish the time of infection. This applies to the diagnosis of Acute T. gondii infection acquired during pregnancy
31
Leishmaniasis IHA

Application: For detection of anti-leishmania antibodies in serum. Interpretation of Results:

1. 2.
The result is reported as follows: If the reported titre is 1:128, send another sample after 2 weeks for re-testing. Significant titres range from 1:256 to 1:12048.
Comments: Low positive titres of 1:32 or more are a sign of Leishmania infection (but does not indicate the status of infection whether active or inactive).
32
Echinococcus IHA
1.
2.
Application: For detection of anti Echinococcus antibodies in human serum. Interpretation of Results: Positive agglutination is given by titres ranging from 1:32 to 1:128. Titres of 1:512 or higher are considered significant for Echinococcosis.
33
Schistosomiasis IHA

Application: For detection of Schistosomia antibodies in serum. Specimen: Serum Interpretation of Result: The result is reported as follows:
1.
2.
If the reported titre is 1:128, may indicate exposure. Titre of patients suffering from schistosomiasis range between 1:256 and 1:1024.
34
Thank you ..
PROF. ABDULKARIM AL-ASKA, FACHARTZ PROFESSOR & CONSULTANT IN INFECTIOUS DISEASES INFECTIOUS DISEASES UNIT DEPARTMENT OF MEDICINE KING KHALID UNIVERSITY HOSPITAL
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Interpretation Of Serology Tests In Selected Infections

Prof. Abdulkarim AlAska

IgM Acute infection IgM cannot cross the placenta