Related Experiment Videos
[DNA typing for HLA-A, B antigens by polymerase chain reaction with sequence-specific primers and clinical
Objective:
To establish DNA typing for HLA-A, B antigens in Chinese by polymerase chain reaction with sequence-specific primers (PCR-SSP).
Methods:
DNA samples were obtained from 178 unrelated donors and 167 kidney recipients. An additional panel of 62 standard DNAs that were typed by UCLA tissue typing lab in USA. A rapid genotyping for HLA-I class (A, B antigens) by PCR-SSP was set up by designed and synthesized 81 specific primers and 1 pair of internal control primer, combining in 61 one-step reactions (20 PCR reactions for A alleles, 41 PCR reactions for B alleles).
Results:
HLA-A, B alleles were successfully typed in 345 clinical samples and 62 standard DNAs by PCR-SSP technique. No false positive or false negative typing results were obtained. Reproducibility was 100% in 40 samples. The overall time of DNA typing was 5 hours. The typing results were consistent with those of UCLA tissue typing lab. Nineteen alleles of HLA-A and 41 HLA-B alleles were accurately distinguished. Thirteen HLA-A alleles and thirty-two HLA-B alleles in Chinese were practically typed.
Conclusion:
DNA typing for HLA-I class (A, B antigens) by PCR-SSP has proved to be a technique of high-resolution, high-specificity, well-reproducibility, and more suitable for clinical application than serology.