Antigen Antibody Reaction

Dr. Mejbah Uddin Ahmed

Antigen Antibody Reaction

Serum:

Fluid that remains after blood

has clotted and cells have been removed.

Antiserum:

Serum containing

antibodies to a specific antigen(s).

Antigen Antibody Reaction

Immunological

test:

Detection

of

antibodies or antigens in any body fluids including serum or tissues.

Serological test: Detection of antibodies or

antigens in serum.

Antigen Antibody Reaction

Titer:

The term titer refers to the highest

dilution of serum at which agglutination still takes place and beyond which no agglutination occurs.

Serology: Study of serum in the context

of antigen or antibody.

Antigen Antibody Reaction

Uses of serologic test:
A. Diagnosis of infectious diseases:

When organism can not be cultured e.g Syphilis. When culture is dangerous! rickettsial disease. When culture techni"ue is not readily available! #$%.

Antigen Antibody Reaction

When culture re"uire too long time! &ycobacteria.

B. Diagnosis of Autoimmune disease: 'b against normal body

components.

. Determination of blood grou!ing and "#A ty!ing.

Antigen Antibody Reaction

Ag$Ab tests are broadly classified into follo%ing ty!es:

(. 'gglutination test ). *recipitation test +. ,omplement fi-ation test (,FT) .. $mmunoassay using labeled reagents /. $mmunofluorescence ($F) 0. 1n2yme linked assay 3. 4adio immunoassay (4$')

Reaction
Agglutination test

Involve particulate antigens and antibodies. Antigens may be: On a cell or attached to latex particles.
Figure 18.4

Antigen Antibody Reaction

Different ty!es of agglutination tests:

Direct: When particulate 'g directly reacts

with the 'b. A.Slide agglutination e.g. 5lood grouping Serotyping of bacteria 5. Tube agglutination e.g. Widal test Weil6Felitest.

Agglutination Reactions
Indirect or !assi&e:
Agglutination of

an antigen, which is not particulate by itself but is coated in carrier particles to make it particulate: R !, "atex, #elatin and $rotein% A of &taph. Aureus.

Agglutination Reactions

Accordingly the named as follo%s :

tests

are

A.'emagglutination e.g. $regnancy test, ($'A, Anti%' bsAg test, ."atex agglutination e.g. ' sAg test, A&O titer. !.$article agglutination e.g. Anti%'I)Ab. *.!o%agglutination e.g. *etection of antigen from !&+ in case of bacterial meningitis

oombs test

Coombs test: Also known as anti% immunoglobulin test because it employs antibodies against immunoglobulin.

A.Direct: (o detect Rh antibody already bound to the surface Rh antigen of fetal R !. B.Indirect: (o detect circulating Rh Ab in the serum of Rh%negative mother bearing the Rh% positive baby.

oombs test
7Direct ,oombs Test 8 Detects antibodies on erythrocytes

+
Patients RBCs Coombs Reagent (Antiglobulin)

oombs test

$ndirect ,oombs Test Detects anti6erythrocyte antibodies in serum

Step 1 Patients Serum Step 2

+
Target RBCs

Coombs Reagent (Antiglobulin)

Reactions
Involve soluble antigens with antibodies

Figure 18.3

tion
'reci!itation '. Single diffusion. in gel:
5. Double diffusion.

'reci!itation in electro!horesis : A. Immune electro!horesis: used for the

diagnosis of paraproteinemia immune deficiency etc.

. ounter Immunoelectro!horesis:
detection of cryptococcus carcinoembryonic antigen etc.

current

used for the antigen in ,SF

tion
Ab in gel

7 &ethod 8 'b in gel 8 'g in a well

Ag

Ag

Ag

Ag

Interpretation: *iameter of ring is proportional to the concentration

Diameter2

Ag Concentration

Immunoelectroph oresis
&ethod 'gs are separated by electrophoresis 'b is placed in trough cut in the agar +
Ag Ag Ab

Ag

Ab

ountercurrent electro!horesis

&ethod 'g and 'b migrate toward each other by electrophoresis 9sed only when 'g and 'b have opposite charges + Ag Ab

om!lement fi(ation Test

Ste!s of

)T:

Ag , Ab- one is known. are mixed. /easured amount of complement is added. If Ag , Ab match , complements will be used. &ensiti0ed R !- R ! 1 anti R ! antibody. is added.

om!lement fi(ation Test

Ag

No Ag

Ag Ag

Patients serum

om!lement fi(ation Test

Interpretation:

*resence of hemolysis indicates that antigen and antibody did not form a comple- i.e test is negative.

'bsence of hemolysis indicates 'g6'b comple- comple- was formed and utili2ed most or all of the complement i.e. test is positive.

om!lement fi(ation Test

!omplement +ixation

Figure 18.9.1

om!lement fi(ation Test

!omplement +ixation

Figure 18.9.2

Reactions
2liminate the harmful effect of a virus or exotoxin

Figure 18.8b

Immunofloures cence
Immunoflourescence: $s highly sensitive but re"uires a special and costly instrument not affordable by small laboratories.

,n+yme$#in-ed Immunosorbent Assay

,#ISA:

Is

very

sensitive,

technically demanding and time consuming but without radiation ha0ards. Once it was very popular and still used for diagnosis of 'I), ' ) and '!) etc.

Radioimmuno assay

RIA: Is highly sensitive, but technically demanding, with possible radiation ha0ards, costly and time consuming. It is used for detection of very small amounts of antigens like hormones, drugs etc.

ommonly Used Immunological Test

Diagnosis of bacterial diseases: A&O titer%$ost streptococcal disease. 3idal test% 2nteric fever. )*R"% &yphilis. ($'A%&yphilis. 3eil%+eix test.

ommonly Used Immunological Test

Diagnosis of bacterial diseases: ' sAg Ati%' sAb Anti%' cAb ' eAg Anti%' eAb Anti%'!)Ab

(hank 4ou