SEROLOGICAL DIAGNOSIS OF VIRUS

INFECTION

PRSENTED BY - SHARMISTHA DAS

REG.NO - 211705350005
BRANCH – MSC.CMB
CONTENT

 INTRODUCTION
 TYPES OF SPECIMEN
 TYPES OF ANTIGEN- ANTIBODY REACTION
 IMMUNOFLUROSCENCE
 COUNTER- IMMUNOELECTROPHORESIS
 WESTERN BLOTTING
 HEMAGGLUTINATION
 RIA
 ELISA
 NEUTRALIZATION
 COMPLEMENT FIXATION TEST
 REFERENCES
INTRODUCTION

 It is the diagnosis of infectious diseases based upon the Antigen-Antibody reactions.

 The type of such reaction depends upon the nature of both Ag and Ab used.
 These reactions are mostly used in the laboratory (In Vitro) to detect either the Ag (Direct
diagnosis) or the Ab (indirect diagnosis).
 PURPOSE :

I. Useful for viruses that grow slowly making recovery in culture media.
II. CMV & hepatitis B virus ( surface antigen) in blood specimens.
III. P24 antigen detection for HIV.
IV. Allowing greater flexibility in the handling & transport of specimens.
TYPES OF SPECIMEN

SYSTEM FOR ISOLATION FOR SEROLOGY

Respiratory system: Throat swab, throat aspirates Paired sera

nasopharyngeal

Central nervous system Feces, blood, CSF Paired sera

Cardiovascular system Feces Paired sera

Skin throat swab , crust, ulcer Paired sera

scrapings & vesicular fluid

Eye Conjuctival scrapings or swab Paired sera

Liver Blood(yellow fever) serum

 Immunofluorescence Test
(IF).
Counter-
immunoelectrophoresis.
Western-blotting.

Hemagglutination.
TYPES OF Ag-
Ab REACTION
Radio immunoassay (RIA).
Enzyme linked
immunosorbent assay
(ELISA).
Neutralization.

Complement fixation Test

(CFT).
 IMMUNOFLUORESCENCE TEST (IF)

 It is Antigen-Antibody reaction where the Antibody is labeled with

fluorescent dye.
 The fluorescent dye is either Fluorescin or Rhodamine linked to
isothiocyanate (ITC).

 TYPE:
I. DIRECT METHOD : to detect specific Ag by the mean of labeling
specific Ab.
II. INDIRECT METHOD : unlabeled Antibody and fluorescently labeled
Anti-Antibody which is better technique to only label one Anti-
Antibody and not all specific Abs to plenty of Antigens needed to
be detected.

 APPLICATION:
I. Helps to identify Rabies virus, Herpes virus, H.Influenzae.
II. detection of immune complexes in biopsy material and detection
of circulating immune cells.
COUNTERIMMUNOELECTROPHORESIS

 Counterimmunoelectrophoresis Test (CIET) is another in vitro

technique for titrating RABV antibodies.

 It is of the binding of an antibody to its antigen with the

addition of an applied electrical field across the diffusion
medium, usually an agar or polyacrylamide gel.

 APPLICATION:
I. It is a rapid and a highly specific method for detection of
both antigen and antibodies in the diagnosis of infectious
diseases including bacterial, viral, fungal, and parasitic.

II. It is commonly used for Hepatitis B surface antigen (HBsAg),

fetoprotein, hydatid and amoebic antigens in the serum, and
cryptococcal antigen in the CSF.
WESTERN BLOTTING
 Western blotting is analysis and detection of proteins was
first developed in 1979.

 Based on the principles of immunochromato- graphy

where denaturated proteins are separated into poly
acrylamide gel according to the isoelectric point and
molecular weight by electrophoresis.

 Then these separated proteins are transferred on a

nitrocellulose or Polyvinylidene fluoride membrane.

 APPLICATION :
I. WB technique are the current confirmatory diagnosis of
HIV in laboratories.
II. Demonstration of specific antibodies in the serum for
diagnosis of neurocysticercosis and tubercular meningitis.
HEMAGGLUTINATION

 It is reaction of viral hemagglutinins with red blood cells

results in a lattice of agglutinated cells that settle irregularly in
a tube or microtiter well, a process known as
hemagglutination.

 APPLICATION:
I. Widely used for the diagnosis of infection caused by
orthomyxoviruses (influenza), paramyxoviruses (measles,
mumps), mononucleosis, arboviruses-togaviruses (including
rubella).
II. The presence of the virus in infected cell cultures can be
detected by hemagglutination.
III. influenza viruses can be detected by hemadsorption test.
RADIOIMMUNOASSAY
 Radioimmunoassay is one of the sensitive immunoassay
techniques which helps in the determination of antigens or
antibodies in a sample with the use of radioisotopes.

 It is an in vitro type of antigen-antibody interaction.

 It was first described in 1960 for the measurement of

endogenous plasma insulin by Solomon Berson and Rosalyn
Yalow.

 APPLICATION :

I. It was first used for the detection of peptide hormones.

II. Detection of different viral antigens.
III. Detection of Hepatitis B surface antigens.
IV. Detection of mycotoxins.
V. Detection of the early stage of cancer.
ENZYME-LINKED IMUUNOSORBENT ASSAY (ELISA)
 Enzyme-Linked Immunosorbent Assay (ELISA) is a modern
molecular technique for the detection of antigen-antibody
interaction with the help of an enzyme.
 The ELISA technique was first conceptualized and developed by
Peter Perlmann and Eva Engvall.

 Types :
I. Indirect ELISA – Antigen is coated to the microtiter well
II. Sandwich ELISA – Antibody is coated on the microtiter well
III. Competitive ELISA – Microtiter well which is antigen-coated is
filled with the antigen-antibody mixture.

 APPLICATION :
I. It is one of the most sensitive and effective methods for the
detection of different viral, bacterial, and fungal infections.
II. Screening test for HIV infection.
III. For the detection of dengue fever, TB, and Hepatitis B infection.
NEUTRALIZATION

 Neutralization is one of the Antigen-Antibody reactions which

aids in reducing the effects of viral antigens or viruses and
toxins produced by different disease-causing microbes such as
bacterial exotoxins.
 The neutralizing antibodies used in virus neutralization tests
can detect the virus along with its various strains
 APPLICATION :
I. Anti- Streptolysin O titre (ASOT) for the diagnosis of post-
Streptococal Complications e.g. Rheumatic fever.

II. Covid-19 neutralizing antibodies test is used to detect the

respective infection in an individual and its severity.

III. It can be used to detect various viral infections such as

measles, mumps, influenza, etc.
COMPLEMENT FIXATION SYSTEM(CFT)

 Complement fixation is one of the most important and

one of the classical techniques for determining antigen-
antibody complexes present in the testing sample.

 The test is based on the principle that the Antigen-

Antibody complex can only fix the complement and its
effect on the hemolysis of RBC used in the indicator
system.

 This is capable of detecting as little as 0.04 mg of

antibody nitrogen and 0.1 mg of antigen.

 APPLICATION : Detection of influenza virus.

REFERENCES

 https://microbenotes.com/complement-fixation-test/

 https://en.m.wikipedia.org/wiki/Radioimmunoassay

 https://www.slideshare.net/MostafaMahmoud76/serological-diagnosis-of-infectious-disease
s

 https://microbenotes.com/neutralization-test-introduction-and-types /
THANK YOU.