ANTIBODY TITRATION

INTRODUCTION
An antibody titer is a measurement of how much antibody an organism has produced that recognizes a particular epitope, expressed as the inverse of the greatest dilution (in a serial dilution) that still gives a positive result. An antibody titration assay should be applied to determine the antibody amount and concentration resulting in the highest signal of the positive population and lowest signal of the negative population. Titers greater >16 are clinically significant.

Source
AIM
The aim of the experiment was to determine the quantity of anti Rh antibody present in a patient serum.
MATERIALS/ EQUIPMENTS
• Test tubes
• Pipette
• Centrifuge
• Rh D positive cells
PROCEDURE
 Label 6 tubes 1-6.
 Place 0.5ml of the patients plasma/serum into tube 1 and 2
 Add 0.5ml of normal saline into tube 2-6.
 Mix thoroughly with a Pasteur pipette
 Pipettel 50ul of the mixture in tube 2 into tube 3.
 Mix thoroughly and pipette 0.5ml of the mixture in tube 3 into tube 4. Do same till tube 6
 Pipette out 0.5ml of the content of tube 6
 Add 25ul of 2.5% cell suspension
 Mix well and incubate at room temperature for 10-15 minutes
 Spin and observe for agglutination/hemolysis in each tube

RESULTS

DISCUSSION
The indirect Coombs test detects the presence of anti-Rh antibodies in a pregnant woman's blood serum. A patient might be reported to have an "indirect Coombs titer" of 16. This means that the patient's serum gives a positive indirect Coombs test at any dilution down to 1/16 (1-part serum to 15 parts diluent). At greater dilutions the indirect Coombs test is negative.
In the results above, Group 1 and 10 gives a titer of 8 and as such all dilution down to 1/8 gives a positive reaction whiles dilutions above give a negative reaction.
Group 2, 3, 7, 9 has a titer of 16 and dilutions down to 1/16 five a positive reaction with different grades of sensitivity
Group 4 and 5 has a higher title of 4 and as such dilutions greater than 1/4 gives a negative test.
Reactions were observed for all dilutions in group 6 and 8. As such the dilution should be continued to determine the titer for which there will be no reaction. A greater dilution of titre indicates more anti Rh antibody present in the patient’s serum whiles a lower dilution indicates less anti Rh antibody.

CONCLUSION
For the results above Groups 6, 7 are clinically significant and should be invested more.

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