Results, Discussion, Conclusion And Standardization Of AHG
After the preparation of the reagent, I carried out a coombs test to affirm if the prepared reagent is actually working. A commercially prepared AHG was used as a control and the test was run in parallel on a prepared sensitized cells. There was agglutination for both reagents when viewed under the microscope indicating that the locally prepared AHG is a working reagent.
1. I prepared sensitized cells
2. Double dilution of the reagent was done using normal saline in the ratio of
3. I then took 6 clean test tubes and labeled one undiluted AHG and the rest according to the dilution made respectively.
4. Then add 1 drop of the sensitized cell in each of the test tubes respectively.
5. Then add 2 drops of undiluted AHG into the tube labeled undiluted AHG and 1 drop each of diluted AHG into the other tubes according to their dilutions.
Standardization of AHG
|Dilution of AHG||Undiluted AHG|
|No of tubes||1||2||3||4||5||6|
|Add sensitized cells||1||1||1||1||1||1 drops|
|Add AHG according to their dilutions||2 drops of undiluted AHG||1 drops||1 drops||1 drops||1 drops||1 drops|
Incubate at 37C for 30mins and then centrifuge for 30 sec. at 1000rpm
Therefore the standardization which is the titre point is
DISCUSSION, CONCLUSION, RECOMMENDATION AND LIMITATIONS
In accurate diagnosis of alloimmune haemolysis, autoimmune haemolysis and drug-Induced immune-mediated haemolysis, the use of AHG is mandatory. This is because AHG is use in the detection of complete and incomplete antibodies in blood transfusion science. The present study includes a detailed procedure carried out on a rabbit as on how to locally produce and standardize AHG.
After the research work, it was discovered that the locally prepared AHG was a successful one with a titre of 1/32. The rabbit used in the research was immunized using the globulin extracted from the human O serum and the sample was collected from the rabbit through heart puncture.
Few comprehensive studies have been carried out in Bangladesh (Rahman et al., 1977) and were successful with a titre of 1/1024 and also in Iran with a titre of 1/4096 (Zandieh et al., 1986). The variance in the titre might be as a result of different breed of rabbit and environment.
Since the local production and standardization of AHG was a successful venture, it can serve as an avenue to bring out the zeal in us as a scientist to embark on research on how to prepare other reagents.
The state and Federal government have to create an avenue both financially and management for medical laboratory scientist to produce reagents locally and not to depend solely on commercially produced reagents.
- In the course of sample collection, heart puncture was used instead of the recommended vein of the ear lobe since it was very small.
- Rabbit of about 10 weeks old where to be used in this research work but since they are difficult to maintain and it was discovered that they cannot withstand the 10 times injections, rabbit of about 4 months old was used.