A new HLA-DRB1 genotyping method using single nucleotide polymorphism (SNP) analysis with multiplex primer extension reactions and its application to mixed samples

Kiyomi Imabayashi, Yuji Yamamoto, Sachiyo Inagaki, Yusuke Doi, Kei Yoshitome, Satoru Miyaishi, Hideo Ishizu

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We have improved on conventional methods for HLA-DRB1 genotyping and devised a new method that is simple, cost-effective, and adequately applicable to routine forensic practice. This method consists of group-specific polymerase chain reaction (PCR) of the exon 2 region of the HLA-DRB1 gene and simultaneous detection of single nucleotide polymorphisms (SNPs) at multiple sites using multiplex primer extension reactions. With this method, we successfully detected HLA-DRB1 genotypes from the following materials: the peripheral blood of 142 donors, 6 aged saliva stains of known DRB1 genotype stored for 5-10 years at room temperature, 10 aged bloodstains of unknown DRB1 genotype stored for 29 years at room temperature, and minimal bloodstains and saliva stains from 3 donors of known DRB1 genotypes. Furthermore, we were able to type DRB1 alleles of the minor component in mixed samples at a proportion of 1/1,000 or 1/10,000. In a criminal case, DRB1 alleles detected from mixed bloodstains on a sword found at the scene enabled us to explain the case. This method is expected to be useful for forensic medicine. Copyright

Original languageEnglish
Pages (from-to)179-194
Number of pages16
JournalActa medica Okayama
Volume59
Issue number5
Publication statusPublished - Oct 2005

Keywords

  • Application to mixed samples
  • Group specific primer
  • HLA-DRB1 genotyping
  • Multiplex primer extension reactions
  • Single nucleotide polymorphism

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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