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

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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.