[Comparative studies of serological typing and HLA-A, B antigen genotyping with PCR using sequence-specific primers]

Di Yi Jun Yi Da Xue Xue Bao. 2004 Nov;24(11):1267-70.
[Article in Chinese]

Abstract

Objective: To evaluate the accuracy of PCR with sequence-specific primers (PCR-SSP) for HLA-I genotyping and analyze the causes of the errors occurring in the genotyping.

Methods: DNA samples and were obtained from 34 clinical patients, and serological typing with monoclonal antibody (mAb) and HLA-A and, B antigen genotyping with PCR-SSP were performed.

Results: HLA-A and, B alleles were successfully typed in 34 clinical samples by mAb and PCR-SSP. No false positive or false negative results were found, and the erroneous and missed diagnosis rates were obviously higher in serological detection, being 23.5% for HLA-A and 26.5% for HLA-B. Error or confusion was more likely to occur in the antigens of A2 and A68, A32 and A33, B5, B60 and B61.

Conclusions: DNA typing for HLA-I class (A, B antigens) by PCR-SSP has high resolution, high specificity, and good reproducibility, which is more suitable for clinical application than serological typing. PCR-SSP may accurately detect the alleles that are easily missed or mistaken in serological typing.

Publication types

  • Comparative Study
  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Genotype
  • HLA-A Antigens / classification
  • HLA-A Antigens / genetics*
  • HLA-A Antigens / immunology
  • HLA-B Antigens / classification
  • HLA-B Antigens / genetics*
  • HLA-B Antigens / immunology
  • Histocompatibility Testing / methods*
  • Humans
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single-Stranded Conformational
  • Serologic Tests*

Substances

  • Antibodies, Monoclonal
  • HLA-A Antigens
  • HLA-B Antigens