Requirements:
- Microscope.
- Normal Saline.
- Bovine Albumin.
- Anti-Human Globulin (Coomb,s Reagent).
- Test tube.
- Centrifuge machine.
- Incubator.
- “O” Group Blood/ “O” Cell Suspension.
Procedure:
- Take 2 drops of “O” cell suspension in a clean test tube.
- Mix 4 drops of direct patient serum in the test tube.
- Centrifuge this test tube at 3400 rpm in the centrifuge for about 15 seconds.
- After centrifugation observe any agglutination in the test tube.
- Mix 2 drops of Bovine Albumin with this solution and incubates this solution 30 minutes at 37°C Temperature.
- After incubation again centrifuge for 5 minutes.
- Take the test tube and discard the supernatant from solution.
- Wash 3 times the remaining sediment/cell with normal saline.
- After washing the sediment add 2 drops of Anti-Human Globulin (AHG) with this suspension.
- Centrifuge it again for 15 Seconds.
- Observe the agglutination in macroscopically as well as microscopically.
- If any agglutination is seen, than Dilute the serum as 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128 with normal saline and repeat the method described above.
- In which step the agglutination will be negative, in the before stage will be result.
- Suppose, the agglutination is negative at the stage of dilution in 1:64 than result will be 1:32.
Note: “O” cell suspension is made from “O”
(+ve) Blood by the washing with normal saline for 3 times. At the last washing
discard all supernatant, keep only the cell in the bottom of the test tube and
mix about 200 micro-liter normal saline.
Comment: This method is tested by me and M-phil
Doctor. I think this is the easy way for Rh-antibody Titre test. Any other
method can be applied but this method is recommended by the Hematologist.
Report sample:
Rh(Anti-D) Antibody: Not Detected
or Rh(Anti-D) Antibody: Detected
Titre: 1:2 or 1:4 or 1:8