IMMUNOLOGY AND SEROLOGY

LABORATORY TECHNIQUES FOR HEPATITIS A
• Total Anti-HAV Antibodies: Competitive Immunoassay
• ELISA
• RIA
- presence of IgG Ab indicates previous
infection
- presence of IgM anti HAV indicates a
recent infection
3 particles present in the sera of infected patients, reactive with anti-HBs Ab
 Dane particle
- the intact infectious B virion
 Spherical noninfectious particles without the viral genome
 Tubular form that is also noninfectious
LABORATORY TECHNIQUES FOR Hepatitis B
• Immunofluorecence
• Rapid one-step immunodiagnostic assay
- uses latex agglutination, cassettes,
membrane test strips or dipsticks
Ex: Equipar , ACON
• Using Recombinant HBsAg to detect Anti-HBs Ab
- an automated enzyme immunoassay
• Detection of HBsAg using a modified Sandwich Assay
- detection: the fluorescence is measured in the
optical assembly, which is a component of the
automated instrument
HEPATITIS C
• Hepatitis C virus is a causative agent of non A and non B hepatitis
• Incubation period is 2- 5 months
• No evidence for transmission in breast milk
LABORATORY TESTS
 Serological Tests based on enzyme immunoassay technology
 Western blot (immunoblotting)
 Tests based on molecular technology which assess viral load, or identify the HCV clade
CYTOMEGALOVIRUS
• Hybrid Capture Technique for detecting CMV-DNA
- based on signal amplification following
nucleic acid hybridization of CMV DNA to
a complementary RNA oligonucleotide
probe. The anti-hybrid Ab that are
affixed to a solid phase capture the hybrids.
• HCT is also available for diagnosis of herpes simplex virus, Chlamydia trachomatis, N. gonorrhoeae
EPSTEIN BARR VIRUS TESTS
 Paul-Bunnell Davidson Differential Test for heterophile Ab (tube method)
 Rapid Test for heterophile Ab (uses cards or slides)
- ex: Equipar, Seradyn Color Slide II Mononucleosis Test
Influenza Virus Tests
 Detection of Anti-viral Ab
• Complement Fixation
• Hemagglutination Inhibition
• Single Radial Immunodiffusion
 Detection of influenza A Ag (“point of care tests”)
• Endogenous viral encoded enzyme assay
• Optical immunoassay

POLIO
3 Factors contribute to the outcome of infection:
1. Virulence of polio strain
2. number of infecting virions
3. immunological status of individual
Laboratory Diagnosis
 Complement Fixation
 Hemagglutinationinhibition
Assays used to measure Anti-HIV Antibodies
 Calypte HIV-1 Urine EIA Test
 Seradyn Sentinel HIV-1 Urine EIA
- uses urine sample
 Home Access HIV Test System
- uses dried blood spot (telemedicine)
 OraSure HIV -1 Western Blot
- uses OMT (oral mucosal transudate)fluid
 Novapath HIV-1 Immunoblot
- western blot; serum/plasma
 HIV-1/HIV-2 ELISA Test Kit
- ELISA/ serum/plasma
 Acon HIV-1/HIV-2
- Chromatographic immunoassay;serum
 Cobas Core Anti-HIV-1/HIV-2-EIA
- Double antigen sandwich assay EIA;
serum/plasma
 Cambridge Biotech HIV-1
- Western blot; urine, serum ,plasma
 AxSYM HIV-1/HIV 2
- automated EIA; serum/plasma
- Western blot; urine, serum ,plasma
Assays that measures Anti-HIV Viral Load
 Amplicor HIV-1 Monitor
-Amplification (cDNA, PCR); plasma
 HIV-1QT
-Amplification (RNA); plasma
 HIV-1 RNA
-Signal Amplification (bDNA); plasma
Test for Aspergillus Fumigatus
• Reverse Agglutination for serum aspergillus-uses latex particles that are coated with anti-aspergillosis galactomannan
• Sandwich Technique- detects Aspergillus Ag in patients serum
• Double Immunodiffusion Assay or Ouchterlony method- screen the presence of anti-aspergillus Ab in serum.
Tests for Cryptococcus neoformans
• Reverse Agglutination- used to indicate the Antibodies that are affixed to the particles( rather than Ag)
• Cryto-LA Test- rapid latex agglutination test that detects the polysaccharide Ag of Cryptococcus in serum or CSF
• Murex Cryptococcus Test- detects cryptococcal in CSF; rapid test (5 min.)

Test for Cryptococcus neoformans
• YA-Crypto Tube Agglutination
-is a tube Antigen system
-detects Antibodies
Tests for Candida Albicans
• Culture Media
Chromogenic medium
Oricult-N-System
-a culture system done for 48 hours at 37 degrees celcius or at room temperature for 5 days.
• Latex Agglutination Test (CANO-TEC)- uses rabbit anti-candida Ag coated beads to agglutinate candida
• Quik-Tri/CAN- reverse agglutination
- is a dual latex agglutination test that can differentiate candida from trichomonas.
BACTERIAL INFECTIONS IN SEXUALLY TRANSMITTED DISEASES
 Venereal Syphilis: Treponema Pallidum
LABORATORY DIAGNOSIS
Serological tests for the diagnosis of syphilis are based on antigen/antibody interactions.
Two types of antibodies produced during infection with T. pallidum.
 anti-lipoidal Ab and anti-T. pallidum antibodies
Non Treponemal Tests
 Are screening procedures for diagnosis of syphilis and are used to detect antibodies to lipoidal materials.
 These tests may not be useful until one to four weeks after the appearance of a chancre and this may give rise to false negative results because the antibody titer is not sufficiently high.

Standard Non Treponemal Tests
 Rapid Reagin Plasma Test
 Is a non treponemal screening test for syphilis and is recommended for screening plasma or serum samples for reagin antibodies.

Venereal Disease Laboratory Test
 A non treponemal slide test in which flocculation is the endpoint.
 The patient’s serum is heated.
 Used to detect antibodies in cerebral spinal fluid specimens to detect neurosyphilis.
Standard Treponemal Test
 Confirmatory test for the diagnosis of syphilis.
 Measures the presence of antibodies to T. pallidum antigen
2 types:
 Fluorescent treponemal antibody absorbed (FTA-ABS)
 T. pallidum immobilization test (TPI)
TESTS
 Second Generation FTA-ABS Test
 Treponema Pallidum Hemagglutination Assay (Confirmatory Test)
 Modified TPHA (Fast latex test)
Non Standard Non Treponemal and Treponemal Test
 Enzyme-Linked Immonosorbent Assay
- Used for both infectious and non-infectious diseases
Syphilis Rapid Test Device
 Rapid qualitative chromatographic immunoassay that uses the affinity of protein A for IgG antibodies to test for treponemal antibodies
GONORRHEA: NEISSERIA GONORRHOEAE
 Laboratory Technique
 Ligase Chain Reaction
-Is a nucleic acid amplification assay based on the ligase chain reaction.
GENITAL TRACT INFECTION:CHLAMYDIA TRACHOMATIS
 Laboratory Diagnosis for Antigen Detection
 Optical Immunoassay
 Mainstream Immunoassay
 Nucleic Acid Amplification
Upper Respiratory Tract Bacterial Infections
 Atypical Pneumonia: Mycoplasma Pneumoniae
 Laboratory Diagnosis
 Tests that detect M. Pneumoniae
a. Specific antigentest
b. Amplification test ( Polymerase
Chain Reaction)
c. Culture
Detecting Antibodies
 Compliment Fixation
 Cold Agglutinin Antibody Titer
 Indirect Immunofluorescence

STREPTOCOCCAL INFECTIONS: STREPTOCOCCACEAE
 Laboratory Diagnosis
a. Antigen Detection: Antibodies or Nucleic
Acid Probes
b. Optical Immunoassay
c. Q test Rapid Antigen Group A Strep
Detection (Antibody Coated Liposomes)
d. Gen-Probe Group A Streptococcal
Direct Test
Serological Tests
 Are used to detect the presence of antibodies specific for streptococcal antigens.
 Antigens used for antibody detection include streptolysin O, DNAse B, Hyaluronidase and Streptokinase
 Sera Test Aso (S-O)
 Commercial kits (DNAse B)
 Wampole (Streptonase B titer test)-for anti DNAse antibodies
WHOOPING COUGH: BORDETELLA PERTUSSIS
 Laboratory Diagnosis
Antigen Detection Technique
-Uses direct fluorescent antibody
staining or nucleic acid amplification tech.,
the polymerase chain reaction,
followed by nucleic acid hybridization for
detection of amplicons, (more sensitive than DFA)
TESTS
 ELISA
Detects anti-pertussis toxin IgG or IgA antibodies.
 Passive Hemagglutination
Potential technique fot widespread screening
LYME DISEASE: BORRELIA BURGDORFERI
 Laboratory Diagnosis
Immunochromatograhic Test
Indirect Fluorescent Antibody Assay
Compliment Fixation
Enzyme Immunoassays
Western Blot (confirmatory test of many infectious agents)
ROCKY MOUNTAIN SPOTTED FEVER: RICKETTSIA RICKETTSII
 LABORATORY DIAGNOSIS
1. Weil-Felix Agglutination Test
2. Latex-R-rickettsii Agglutination test
LEPTOSPIROSIS: LEPTOSPIRA INTERROGANS
 LABORATORY DIAGNOSIS
1. Lateral Flow Immunoassay
2. LEPTO Dri- Dot Agglutination
SALMONELLOSIS AND TYPHOID FEVER: SALMONELLA SPECIE
 LABORATORY DIAGNOSIS
Chromogenic Culture Medium- selective for Salmonella species
Serological Tests
1. DIP-S-TICK Assay (enzyme immunoassay)
2. Widal Agglutination Test (counter immunoelectrophoresis)
MEASLES, MUMPS, RUBELLA AND VARICELLA
 MEASLES
1. ELISA- detects anti-measles IgM
2. Indirect Immunofluorescence Antibody-detects anti-measles antibodies serum.
MUMPS
 Laboratory Diagnosis
Enzyme-Linked Fluorescent Immunoassay (ELFA)
RUBELLA
 LABORATORY DIAGNOSIS
Enzyme Immunoassay for Anti-Rubella Antibodies
Capture Immunoassay for IgM
Non Serological Test
Amplification techniques
VARICELLA
 Laboratory Diagnosis
ELISA