[DNA typing for HLA-A by polymerase chain reaction with sequence-specific primers].
Tan J, Xiao Ping Tang
Abstract
Tan J, Xiao Ping Tang
Abstract
The authors have established a method of DNA typing for HLA-A antigen in Chinese by using polymerase chain reaction with sequence-specific primers (PCR-SSP). HLA-A alleles were successfully typed in all clinical samples and 36 standard DNAs by PCR-SSP. A total of 635 A antigens were typed in 345 donor-recipients, including 55 homozygotes and 290 heterozygotes. Twenty alleles of HLA-A were accurately distinguished by DNA typing. No false positive or false negative typing results were obtained. Reproducibility was 100%. The overall time of DNA typing was 5 hours. The typing results were concordant with the results of UCLA Tissue Typing Lab. Comparative study of HLA-A typing was carried out using PCR-SSP and standard serology. The results showed that the discrepancy rate of serology was 9.0%, containing 13 individuals incorrectly interpreted by serology and 16 serological "blanks" turned out to be a definable alleles by PCR-SSP. In conclusion, DNA typing for HLA-A by PCR-SSP has proved to be a high-resolution, high-specificity, rapid and accurate technique, suitable for clinical application with a greater precision than serology.
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The authors have established a method of DNA typing for HLA-A antigen in Chinese by using polymerase chain reaction with sequence-specific primers (PCR-SSP). HLA-A alleles were successfully typed in all clinical samples and 36 standard DNAs by PCR-SSP. A total of 635 A antigens were typed in 345 donor-recipients, including 55 homozygotes and 290 heterozygotes. Twenty alleles of HLA-A were accurately distinguished by DNA typing. No false positive or false negative typing results were obtained. Reproducibility was 100%. The overall time of DNA typing was 5 hours. The typing results were concordant with the results of UCLA Tissue Typing Lab. Comparative study of HLA-A typing was carried out using PCR-SSP and standard serology. The results showed that the discrepancy rate of serology was 9.0%, containing 13 individuals incorrectly interpreted by serology and 16 serological "blanks" turned out to be a definable alleles by PCR-SSP. In conclusion, DNA typing for HLA-A by PCR-SSP has proved to be a high-resolution, high-specificity, rapid and accurate technique, suitable for clinical application with a greater precision than serology.
Key concepts: Typing, Polymerase chain reaction, Serology, Biology, Human leukocyte antigen, Genetics, Molecular biology, Virology