Blood Grouping Technique(Rh Typing)

Slide Method:

1.    Place 3 drops of reagent Anti Rh (D) on a labeled slide
2.     Put one drop of 40-50% red cells suspended in plasma or serum and 1 drop of each D positive and D negative cells on each drop of anti Dreagent.
3.     Mix the cell suspension and reagent separately, using clean stick for each test and spread the mixture evenly on the slide over an area of 15 mm diameter.
4.    See the agglutination after 2 minutes.


Tube method:

1.    Label3 tubes as test, pos. control and negative control
2.    Place 1 drop of anti Dreagent to each tube
3.     Add 1 drop of 2-5% washed cell suspension in saline to the tube “test and known RhD positive and negative cells to tube positive control & negative control.
4.    Incubate the tubes at 37°C for 10 min. and centrifuge at 1000 rpm for 1 minute.
5.     Gently resuspend the button and observe for agglutination, all negative results must be confirmed under microscope



.
Interpretation:

 A positive test has agglutination with anti-Rh (D) in test cells and positive control cells and smooth suspension of the cells in the negative control.
A negative test has a smooth suspension of cells in the test cells and negative control and agglutination in positive control.




Test
( Unknown Sample)
Positive control
( Known Rh positive cells )
Negative Control
( Known Rh Negative control)
Result
+
+
O
Rh(D) positive
O
+
O
Rh( D)
negative


                                     +         =         Agglutination   O  =   no agglutination

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