All about Passive agglutination
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PASSIVE AGGLUTINATIONGROUP-K
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• What is passive agglutination?• What are various types of passive agglutination tests?
AREAS OF INTERESTS…
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LET’S GET STARTED…
What is agglutination?
Particulate antigen + its specific antibody
Electrolytes at an optimal temperature and pHVisible clumping of
particles
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LET’S GET STARTED…
What is precipitation
reaction?
Soluble antigen + its specific antibody
Electrolytes at an optimal temperature and pH
Insoluble precipitate
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LET’S GET STARTED…
And what is Passive Agglutination??
Precipitation reaction
Agglutination testsco
nvert
ed
By attaching soluble antigens to the surface of carrier particles such as latex particles, bentonite, RBCs, etc.
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ADVANTAGES OF PASSIVE AGGLUTINATIONAdvantage of Passive agglutination over precipitation tests are:• More convenient• More sensitive for detection
of antibodies• More sensitive for detection
of antigens (Reverse passive)
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REVERSE IS POSSIBLE…
When instead of antigen, the antibody is adsorbed on the carrier particles for estimation of antigens, it is known as Reverse passive agglutination
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LATEX(0.8-1µ)
ANTIBODY
LATEX
LATEX
LATEX
LATEXANTIGEN (in patient
serum)
LATTICE FORMATION
REVERSE PASSIVE AGGLUTINATION…
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Simplifying the course of study...
Passive agglutination
Coagglutination test
Latex agglutination test
Hemagglutination test
On basis of carrier particle used
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COAGGLUTINATION TEST
Staphylococcus aureus (Cowan 1 strain) used
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COAGGLUTINATION TEST
S.Aureus (Cowan 1)
Fc
FabFab
ANTIBODY (IgG)
ANTIGEN (in patient sreum)
PROTEIN AY
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COAGGLUTINATION TEST
S.Aureus (Cowan 1)
ANTIBODY (IgG)
S.Aureus (Cowan 1)
ANTIGEN (in patient sreum)
LATTICE FORMATION
S.Aureus (Cowan 1)
S.Aureus (Cowan 1)
S.Aureus (Cowan 1)
PROTEIN A
How it works?
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LATEX AGGLUTINATION TEST...
The carrier particle is Latex or polystyrene latexBrains behind this: C M Plotz and J M Singer
Accidently discovered IgG adsorbed naturally to polystyrene latex
particles (1953)
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LATEX AGGLUTINATION TEST...
How it works?LATEX(0.8-1µ)
AntigenLATEX
LATEX
LATEX
LATEX
LATTICE FORMATION
ANTIBODY
ANTIGEN
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LATEX AGGLUTINATION TEST- USES
• Carrier + Antibody- detection of antigens- CRP,RA factor, HCG, Hepatitis B
• Carrier + Antigen- antibodies to meningococci, H.influenzae type b
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Large number of antigens can adsorbed on carrier Better visualization of Ag-Ab reaction due to larger
particle size of Latex beads preventing previous cumbersome process involved in precipitation reactions (no sophisticated equipments required)
Latex particles do not cross-react with other antibodies
Less time consuming
LATEX AGGLUTINATION TEST- ADVANTAGES
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HEMAGGLUTINATION TEST
The carrier particle is Red Blood Cell/Tanned blood cell (Goose RBCs preferred)
Brain behind this: George Hirst (1942)
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HEMAGGLUTINATION TEST...
How it works?RBC
AntigenRBC
LATTICE FORMATION
ANTIBODY
ANTIGEN
RBC
RBC RBC
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HEMAGGLUTINATION TEST- Examplified by Rose Waaler Test (Carrier-RBC )
RBC
Amboceptor(subagglutinating dose)
RBC
LATTICE FORMATION
RA FACTOR (in patient serum)
AMBOCEPTOR (Rabbit anti-sheep erythrocyte antibody)
RBC
RBC RBC
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HEMAGGLUTINATION TEST- Examplified by TPHA (Carrier-Tanned RBC )
TannedRBC
Antigen(T.pallidum extract)
TannedRBC
LATTICE FORMATION
ANTIBODIES against T.pallidum (in patient serum)
ANTIGEN (T.pallidum extract)
TannedRBC
TannedRBC
TannedRBC
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VIRAL HEMAGGLUTINATION AND HEMAGGLUTINATION INHIBITION TEST
What is Viral Hemagglutination?
RBC
VirusRBC
LATTICE FORMATION
VIRUS
RBC
RBC RBC
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VIRAL HEMAGGLUTINATION AND HEMAGGLUTINATION INHIBITION TEST
What is Hemagglutination inhibition ?
VirusNO LATTICE FORMATION
VIRUS
RBCAb against VIRUS
RBCRBC
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AGGLUTINATION TESTS IN OUR LAB IN A BIRD’S EYE VIEW
ASO titre
RA factor detection
CRP
KIT- Syphicheck
Infectious mononucleosis (Immutex)
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LET’S END BY LOOKING AT RECENT ADVANCES
• Determination of anti-streptolysin O antibody titer by a new passive agglutination method using sensitized toraysphere particles.
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THANK YOU
References from:Anantnarayan and
Paniker’s Textbook of microbiology
C.P. Baweja’s Textbook of Microbiology
Subhash Chandra Parija’s Textbook of microbiology and ImmunologyAmerican Society for microbiolgy-Journal of Clinical Microbiology