Blood Grouping Technique(ABO)

Slide Method :  Used only for preliminary group testing of donor .

1.    The slide test may be performed on clean microscopic or ceramic opaque tiles
2.     Put 1 of anti-A and 1 drop of Anti-B separately on the labeled slide or tile.
3.    Add 1 drop ofabout 40-50 % red suspension or 1 drop of whole blood oftest sample to each drop of reagent.
4.    Mix the cell and reagent using a clean stick. Spread each mixture evenly on the slide over an area of 15 mm diameter.
5.    Rock-rotate the slide or plate and leave the test for 2 min. at room temperature and look for agglutination.
6.    Record the result.

Tube method:
1.    Prepare 2-5% cell suspension of test sample in normal saline.
2.    Set up three rows of clean test tubes and label them. Add two volumes(2 drops) of Anti-Ain the tube labeled 'A', 2 drops of Anti-B in tube labeled B and 2 drops of Anti-AB in tube labeled AB.
3.     Add 1 volume (1 drop) of 2-5 % suspension of test sample in each tube.
4.    Mix and incubate at room temperature for 30-45 minutes or spin after 5-10 minutes.
5.    Observe the supernatant fluid for the presence of haemaolysis against a well lighted background.
6.  Gently disperse the cell button and check for agglutination against a well lighted
Background.
7.    Where no agglutination is seen macroscopically, examine the contents under microscope.
8.    Record results immediately.





Serum grouping:
1.   Label three tubes A, B & O
2.   Place 2 drops of test serum in each tube
3.   Add 1 drop of A cells to tube A, 1 drop of B cell to tube B and 1 drop of O cell to tube O
4.   Mix and incubate at room tem. For 30-45 min. or centrifuge after 5-10 minutes.
5.   Observe the supernatant fluid for the presence of haemaolysis
6.   Gently disperse cell button and see for agglutination
7.   All negative results must be examined under microscope.



Result
Reaction of RBC with
Reaction of serum with pooled cells
Blood group of the tested cells
Anti-A
Anti-B
Anti-AB
+
+
+
+
O
+
O
+
+
O
O
O
O
O
O
A
B
O
O
O
O
O
+/H
O
+/H
O
O
+/H
+/H
O
+/H
+/H
+
        

AB
A
B
O
Oh






+ =Agglutination           o =No Agglutination        H=Haemolysis

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