Agglutination:
Is the clumping of antibodyantigen complex to form insoluble and
visible aggregates.
The word agglutination comes from the Latin agglutinare, meaning "to glue to."
Antigen:
Antigen is any molecule that binds specifically to an antibody.
Antibody:
Antibodies (immunoglobulins) are proteins secreted by the immune system
to identify and neutralize foreign objects (antigens), such as bacteria and
viruses.
Zone of equivalence:
Antibodies and antigens are present in an optimum ratio.
This leads to cross-linkages between acells or particles, so
agglutination appers (positive reaction).
Test procedure:
Add 1 drop CRP latex to 1 drop of patient serum (may be diluted)
then stir & mix.
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Sample:
Blood drawn into EDTA is preferred but oxalated or citrated blood may
be used.
The blood sample should be tested as soon as possible after collection
and should not be stored.
Isotonic saline
Pipettes
12 x 75 mm tubes
Centrifuge
Microscope
Procedure:
1. Put 1-2 drops of blood in EDTA tube.
2. Wash this tube three times with isotonic saline.
3. Prepare a 5-10% suspension from the washed cells.
4. Add one drop of the washed 5-10% suspension to a tube.
5. Then, add one drop Polyspecific Antihuman Globulin and shake to mix.
6. Centrifuge the tube for 30 seconds.
7. Immediately resuspend gently and examine for agglutination.
Interpretation:
1. No agglutination indicates a negative DAT - nothing is coating the
patient's cells in vivo.
2. Agglutination with Polyspecific AHG indicates a positive DAT, due to
either IgG antibodies or complement coating the patient's cells.
Preccautions:
AHG was omitted inactive or was diluted out by too much residual
saline in the tube.
Drain the last wash well and blot before adding AHG.
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