DNA typing of the HLA-A gene: population study and identification of four new alleles in Japanese

Tissue Antigens. 1996 Feb;47(2):93-101. doi: 10.1111/j.1399-0039.1996.tb02520.x.

Abstract

With the use of polymerase chain reaction (PCR) and sequence-specific oligonucleotide probe (SSOP), we established a DNA typing method of the HLA-A locus. A pair of primers to amplify the highly polymorphic region of HLA-A gene including exon 2 and exon 3 was designed and the amplified DNAs were hybridized with 91 types of 32P labeled SSOPs. This method allowed discrimination of all known HLA-A alleles except for two combinations, A*0201 or A*0209 and A*0207 or A*0215N, which have identical sequences in exon 2 and exon 3. Another pair of primers was designed for amplification of exon 4 and the PCR products were hybridized with 5 SSOPs to distinguish A*0201 and A*0207 from A*0209 and A*0215N, respectively. In this study, 81 B-lymphoblastoid cell lines (BLCL) homozygous for HLA and 553 unrelated healthy Japanese individuals were determined for their HLA-A genotypes. Based on the genotyping results, frequency of HLA-A alleles and linkage disequilibrium between HLA-A and HLA-B in the Japanese population were investigated. In addition, four new HLA-A alleles were identified and their nucleotide sequences in exon 2 and exon 3 were determined to confirm the typing results.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • B-Lymphocytes / cytology
  • Base Sequence
  • Cell Line
  • DNA
  • DNA Primers
  • Exons
  • HLA-A Antigens / genetics*
  • Humans
  • Japan
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes
  • Polymerase Chain Reaction
  • Population Surveillance
  • Sequence Analysis

Substances

  • DNA Primers
  • HLA-A Antigens
  • Oligonucleotide Probes
  • DNA