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Immunology Practical With Answer

The document outlines various immunological techniques, including conventional methods like agglutination and precipitation, as well as newer methods such as ELISA and immunofluorescence. It details the principles, types, and applications of these methods for detecting antigens and antibodies in clinical samples. Additionally, it discusses point-of-care testing principles and formats for rapid disease diagnosis.

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3 views3 pages

Immunology Practical With Answer

The document outlines various immunological techniques, including conventional methods like agglutination and precipitation, as well as newer methods such as ELISA and immunofluorescence. It details the principles, types, and applications of these methods for detecting antigens and antibodies in clinical samples. Additionally, it discusses point-of-care testing principles and formats for rapid disease diagnosis.

Uploaded by

gharini71
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Immunology practical with answer

Q1

1. Conventional
a. Agglutination
b. Precipitation

2. Newer methods
a. ELISA
b. Immunofluorescence

Q2

Agglutination

1.Principle: When a particulate or insoluble antigen is mixed with its antibody in the presence of
electrolytes at a suitable temperature and pH, the particles are clumped or agglutinated

2. Types

Direct Agglutination Test:

Slide Agglutination - blood grouping and cross matching

Tube Agglutination - Widal test, Brucella (Standard agglutination test), Coombs


antiglobulin test

Indirect Agglutination Test or Passive Agglutination Test (for Antibody Detection)

Latex Agglutination Test (LAT) for Antibody Detection: ASO (antistreptolysin O antibody)

Reverse Passive Agglutination Test (for Antigen Detection)

1. C reactive protein
2. Coagglutination test:

Hemagglutination: Test It refers to the agglutination tests that use RBCs as source of antigen.
Hemagglutination tests are of two types: direct (described below) and indirect (or IHA, obsolete
now).

Direct Hemagglutination Test

Paul Bunnell test:

Cold agglutination test:

Blood grouping (ABO and Rh grouping)

Coombs test or antiglobulin test:

Q3 PRECIPITATION REACTION
When a soluble antigen reacts with its antibody in the presence of optimal temperature, pH and
electrolytes (NaCl), it leads to formation of the antigen–antibody complex in the form of:

Insoluble precipitation band when gel or agar containing medium is used (called immunodiffusion) or

Insoluble floccules when liquid medium is used (called flocculation test). Slide Flocculation Test (for
Syphilis) – RPR

Q4 ELISA

ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) ELISA is an immunoassay that detects either


antigen or antibodies in the specimen, by using enzyme–substrate– chromogen system for
detection
1. Principle of ELISA
 Immunosorbent: Indicates an absorbing material is used (e.g. polystyrene, polyvinyl)
absorbs the antigen or antibody present in serum
 Enzyme: Used to label one of the components of immunoassay (i.e. antigen or
antibody). Substrate-chromogen system: A substrate-chromogen system is added at the
final step of ELISA
 The enzyme reacts with the substrate, which in turn activates the chromogen to produce
a color ™ The classical example is, horseradish peroxidase used as enzyme which reacts
with its substrate (hydrogen peroxide), that in turn activates the chromogen
(tetramethyl benzidine) to produce a color
 The color change is detected by spectrophotometry in an ELISA reader. Intensity of the
color is directly proportional to the amount of the detection molecule (Ag or Ab) present
in test serum.

Types

Direct ELISA

Indirect ELISA

Sandwich ELISA

IgM Antibody Capture (MAC) ELISA

Competitive ELISA

ELISPOT Test

Applications of ELISA ELISA can be used both for antigen and antibody detection.

ELISA used for antigen detection: Hepatitis B [hepatitis B surface antigen (HBsAg) and precore antigen
(HBeAg)], NS1 antigen for dengue, etc.

ELISA can also be used for antibody detection against hepatitis B, hepatitis C, HIV, dengue, EBV, HSV,
toxoplasmosis, leishmaniasis, etc
Q6 Immunofluorescence

Fluorescence refers to absorbing high energy-shorter wavelength ultraviolet light rays by the fluorescent
compounds and in turn emitting visible light rays with a low energy-longer wavelength.

The fluorescent dye is used to conjugate the antibody and such labeled antibody can be used to detect
the antigens or antigen–antibody complexes on the cell surface

The fluorescent compounds commonly used is fluorescein isothiocyanate (FITC).

Types Direct Immunofluorescence Assay

Indirect Immunofluorescence Assay

Applications:

Detection of autoantibodies (e.g. antinuclear antibody) in autoimmune diseases

Detecting microbial antigens, e.g. rabies antigen in corneal smear

Detection of viral antigens in cell lines inoculated with the specimens.

Q8 Point of care

Two principles of rapid tests are available—lateral flow assay and flow through assay

Both the formats are available for the diagnosis of various diseases such as malaria, hepatitis B, hepatitis
C, HIV, leptospirosis, Helicobacter pylori, syphilis

Types:

Immunochromatographic Test

Flow-through Assay

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