Related Experiment Videos
DNA typing system for HLA-A2 alleles by polymerase chain reaction with sequence-specific primers
Objective:
To establish a PCR-SSP method for discriminating as many HLA-A*02 alleles, which could easily be introduced into a routine laboratory.
Methods:
In this study we typed HLA-A*02 polymorphisms by a sequence-specific primer (SSP) method, which involved round 1 and round 2 PCR reactions to detect 17 HLA-A*02 alleles (they are HLA-A*0201-0217 alleles) covering exon 2 and exon 3.
Results:
We have found that DNA sample concentration and purity were the most important variable in determining the quality of the result. For identifying correct band size, the size marker used was important. We noticed that different PCR machines performed differently. By this method, we detected 20 HLA-A*02 positive genomic DNA samples and found 4 kinds of HLA-A*02 alleles. They were HLA-A*0201, 0203, 0206 and 0210.
Conclusion:
The HLA-A*02 PCR-SSP method was proven to be a reliable and easily applicable typing method. Our results suggest that the SSP described here provides an optimal HLA-A*02 typing technique that may be useful in selecting donor-recipient pairs in bone marrow transplantation between unrelated individuals.