Serological Testing
Serological tests are laboratory methods that detect the presence of
antibodies or antigens in a patient's serum to diagnose infections, autoimmune
diseases, and blood compatibility.
II. Principles of Serological Testing
1. Antigen-Antibody Reaction:
Antigen: Foreign substance (e.g., virus, bacteria)
Antibody: Host immune protein (IgM, IgG) produced in response to
antigen
2. Types of Antibodies:
IgM: First antibody produced, indicates recent/acute infection
IgG: Indicates past infection or immunity
3. Detection Methods:
Direct detection of antigens
Indirect detection of antibodies
III. Types of Serological Tests
Test Type Description Example Diseases
Agglutination tests Visible clumping due Widal test, latex
to Ag-Ab binding agglutination
Precipitation tests Insoluble complexes Ouchterlony (double
form and precipitate diffusion)
Complement fixation Detects complement Viral infections, some
usage in antigen- parasites
antibody reaction
ELISA (Enzyme-linked Detects Ab or Ag using HIV, hepatitis B/C,
Immunosorbent Assay) enzyme reaction dengue
Western blot Protein separation + HIV confirmatory,
detection of specific Lyme disease
antibodies
Immunofluorescence Fluorescent-labeled Autoimmune disorders
(IFA) antibodies detect (ANA test)
specific antigens
Radioimmunoassay Uses radiolabeled Hormones, hepatitis B
(RIA) substances surface antigen
Rapid diagnostic tests Lateral flow/strip tests COVID-19, malaria,
(RDTs) for quick results dengue
IV. Common Serological Tests and Their Applications
1. HIV Testing
Screening: ELISA (detects anti-HIV antibodies)
Confirmation: Western blot, PCR
Window period: ~3 weeks post-exposure
2. Hepatitis Panel
HBsAg – Hepatitis B surface antigen → Active infection
Anti-HBs – Immunity (past infection or vaccine)
Anti-HBc IgM/IgG – Recent or past infection
Anti-HCV – Screening for Hepatitis C
3. Syphilis
Non-treponemal: VDRL, RPR (screening)
Treponemal: FTA-ABS, TPHA (confirmation)
4. Dengue
NS1 antigen (early phase)
IgM/IgG detection by ELISA or RDT
5. Typhoid Fever
Widal Test:
Detects agglutinating antibodies (O and H)
Titers ≥1:160 are significant, but interpretation depends on local baseline
6. COVID-19
IgM/IgG tests (detect post-infection)
Antigen tests: detect viral proteins
PCR: confirms presence of viral RNA (not serological)
7. Autoimmune Diseases
ANA (antinuclear antibody) test (IFA method) → Lupus, RA
Anti-dsDNA, Anti-Smith antibodies → SLE
V. Interpretation of Serologic Results
Result Interpretation
IgM positive Current or recent infection
IgG positive, IgM negative Past infection or immunity
Both IgM and IgG positive Recent infection transitioning to
chronic
Negative IgM and IgG No exposure or early window period
VI. Advantages and Limitations
✅ Advantages:
Detect past or current infections
Non-invasive (blood/serum)
Useful in screening and epidemiological surveys
❌ Limitations:
Cross-reactivity (false positives)
Window period (false negatives early in infection)
Requires quality control and clinical correlation
VII. Summary Table: Key Serological Tests
Disease Test(s) Used Target Detected
HIV ELISA, Western blot Anti-HIV antibodies
Dengue NS1, IgM/IgG Antigen, antibodies
Syphilis VDRL, FTA-ABS Non-specific & specific
Ab
Hepatitis B HBsAg, Anti-HBs, Anti- Antigens and
HBc antibodies
COVID-19 IgM/IgG, Antigen, RT- Antigen, antibodies,
PCR RNA
Typhoid Widal test Agglutinating
antibodies
VIII. Clinical Tips
Use ELISA for screening, Western blot/PCR for confirmation
Timing of sample is crucial (early vs late infection)
Always interpret serological tests in context with clinical findings
