Anti Human Globulin Test

Anti Human Globulin Test

Discovered by comb's and Pace
This test is against the human protein mainly Globulin.

Preparation

1human serum is injected in arabbit.

2after 15 days sucked the blood sample form rabbit and purified because the human serum contains IgG and C3(complement factor).

This 15 days are the period of production of IgG against the human serum.

Principle of AHG

Change(convert)the in visible senstization reaction to visible agglutination reaction.
Mechanism
Acts to link between the IgG antibody which coat the RBCs
Coombs Reagent

1Polyspecific reagent

Containing both anti IgG and antiC3
2monospecific reagent

Either containing anti IgG or anti C3
To explain the reaction a monospecific reagent is used 2method
1direct AHG(direct coombs test

Aim:-to detect the invivo sensitization of RBCs
Method

1whol blood sample is centrifuged
2discarde the plasma
3keep the packed cell
4washing the cell by normal salin for 3 time as minimum
5centr ifuge 3500 RPM for 3-5minut
6discarde the super natal except small amount of cell
7Repeat this for 2time as minimum to remove the free Anti body which are found in plasma so as prevent the test false –ve result
8discarde all normal salin and add coombs in the middles of tube
9centrifuge for less than1minute to accelerate the reaction
10read the reaction
Result

Positive agglutination
Negative No agglutination
The pt RBCs are sensitized invivo or the Pt RBCs are coated by IgG
Comment:-
Incomplete anti body
2Indirect AHG(in direct coombs test

Aim:
to detect the invetro sensation of RBCs
Method

1Pt serum is incubated with +ve pooled cells in water path in37c for 45minute
2Apply the direct coombs test
3centrifuge for less than one minute
Result

ve+ RBCs are sensitized invetro
Comment
Pt serum contains IgG or C3 complement

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