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PASSIVE AGGLUTINATIONGROUP-K
• What is passive agglutination?• What are various types of passive agglutination tests?
AREAS OF INTERESTS…
LET’S GET STARTED…
What is agglutination?
Particulate antigen + its specific antibody
Electrolytes at an optimal temperature and pHVisible clumping of
particles
LET’S GET STARTED…
What is precipitation
reaction?
Soluble antigen + its specific antibody
Electrolytes at an optimal temperature and pH
Insoluble precipitate
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.
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)
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
LATEX(0.8-1µ)
ANTIBODY
LATEX
LATEX
LATEX
LATEXANTIGEN (in patient
serum)
LATTICE FORMATION
REVERSE PASSIVE AGGLUTINATION…
Simplifying the course of study...
Passive agglutination
Coagglutination test
Latex agglutination test
Hemagglutination test
On basis of carrier particle used
COAGGLUTINATION TEST
Staphylococcus aureus (Cowan 1 strain) used
COAGGLUTINATION TEST
S.Aureus (Cowan 1)
Fc
FabFab
ANTIBODY (IgG)
ANTIGEN (in patient sreum)
PROTEIN AY
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?
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)
LATEX AGGLUTINATION TEST...
How it works?LATEX(0.8-1µ)
AntigenLATEX
LATEX
LATEX
LATEX
LATTICE FORMATION
ANTIBODY
ANTIGEN
LATEX AGGLUTINATION TEST- USES
• Carrier + Antibody- detection of antigens- CRP,RA factor, HCG, Hepatitis B
• Carrier + Antigen- antibodies to meningococci, H.influenzae type b
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
HEMAGGLUTINATION TEST
The carrier particle is Red Blood Cell/Tanned blood cell (Goose RBCs preferred)
Brain behind this: George Hirst (1942)
HEMAGGLUTINATION TEST...
How it works?RBC
AntigenRBC
LATTICE FORMATION
ANTIBODY
ANTIGEN
RBC
RBC RBC
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
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
VIRAL HEMAGGLUTINATION AND HEMAGGLUTINATION INHIBITION TEST
What is Viral Hemagglutination?
RBC
VirusRBC
LATTICE FORMATION
VIRUS
RBC
RBC RBC
VIRAL HEMAGGLUTINATION AND HEMAGGLUTINATION INHIBITION TEST
What is Hemagglutination inhibition ?
VirusNO LATTICE FORMATION
VIRUS
RBCAb against VIRUS
RBCRBC
AGGLUTINATION TESTS IN OUR LAB IN A BIRD’S EYE VIEW
ASO titre
RA factor detection
CRP
KIT- Syphicheck
Infectious mononucleosis (Immutex)
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.
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