Antigen-Antibody reactions

Difference between preçipitation & agglutination reaction

A. Precipitation nxns
4 1 Ab.

1) Ring test
Eg. Ascoli's thermoprecipitation test for Anthrax, CRP, Lance field
serotyping
2) Slide flocculation test: when a drop of Ag soln and
pt's serum are placed on a slide and
mixed well by shaking, the reaction
appears in form of floccules
Eg. VDRL test for Dx of Syphillis
3) Tube flocculation test
Eg. Kahn test for diagnosis of Syphilis
4) Immunodiffusion test
a. Single diffusion in 1 dimension / Oudin procedure
b. Double diffusion in 1 dimension/ Oakley Fulthorpe procedure
C. Single diffusion in 2 dimension /Mancini method
d. Double diffusion l2 dimension /Ouchterlony's method
e. Elecro-immunodiffusion
i. Immune-electrophoresis
i. Rocket electrophoresis
ii. Counter-current immunoelectrodiffusion

B. Agglutination Rxns
1) Slide agglutination. Eg: Blood grouping, identifiacton of bacterial isolate
2) Tube agglutination. Eg Widal test (enteric fever)

Heterophile agglutination erophle Ag of Kiksksiae cauung hyphe

kais
Weil-felix test -

Rickettsial disease e SAeue by wkn

Paul-Bunnel test - Infectious mononucleosis-¢gv shasezambiienity wih shees
Cold ox RBs.
agglutination test- Mycoplasma pneumoniae
3) Passive agglutination- eg. detection of ASO, CRP, RA factor
A precipitation rxn can be converted into agglutination by coating soluble Ag onto the surface of carrier
molecule (eg.
Latex / Bentonite/ RBBs) (when instead of Ag, Ab coated onto carriermolecule-Reverse Passive
ARglutination)
4) Coomb's test / Anit-globulin test- Direct Coomb's and Indirect Coomb's
YorA
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 REsEARCH
VDRL test VeNCeAL D1SCA SE LABoeA 1OR Y
Developed in VDRL Laboratory in New York
piaced slide
Principle: slide flocculation test (when a of Ag soin and pt's serum
drop are on a

and mixed well by shaking, the reaction appears in form of floccules)

Procedure:
Quantitativetest Qualitative test
Carried out on VDRL tile (with mutiple Serial doubling dilutions are prepared
and used for the test
concavities)
Inactivated drop of serum is added on Highest dilution found reactive is taken

to the concavity as titre

A drop of VDRL Ag is then added and
mixed with stick
Rotated manually on VDRL rotator fixed
at 180 rpm for 4-8 mins

Results read under microscope

Interpretation: Interpretation
Presence of floccules -reactive Reactive
Absence of floccules non-reactive Weakhy reactive
Not reactive
A titre of 1:8 or more significant

Slide Agglutination
Eg. Blood grouping, identifiacton of bacterial isolate
to a smooth uniform suspension of particulate Ag
I n this test agglutination results when
of Anti-serum is added and mixed with
in a drop of saline on a glass slide or tile, drop
a

a wire loop
in a few seconds indicates +ve result
Clumping together of particles and clearing of drop
taken in
control on same slide; for this Ag suspension is
I t is necessary to have negative
a drp of saline without antiserum
it rules out that Ag is not auto-agglutinable
I f no clumping occurs,
Nidal test (for enteric fever)

Principle: tube agglutination test: which detects Abs produced against S.typhi, S. paratyphi A
&B

Antigens used
H &0 of S. typhi
Hof S. paratyphiA
H of S. paratyphi B

Paratyphoid O Ags are not used as they cross-react with typhoid O antigens by sharing factor
12

Procedure
Widal rack with 4 of test tube for 4 Ags is used for the test
rows

Equal volume of serial dilution of serum is added to all test tubes

Antigens are then added to respective rows
Rack is incubated @ 37°C
Control tubes containing NS and Ags are also used to check auto-agglutination

Interpretation:
H agglutination- loose cotton wooly clumps
agglutination- granular disc-like pattern at bottom of test tube