Haplotyping the human leukocyte antigen system from single chromosomes

Sci Rep. 2016 Jul 27:6:30381. doi: 10.1038/srep30381.

Abstract

We describe a method for determining the parental HLA haplotypes of a single individual without recourse to conventional segregation genetics. Blood samples were cultured to identify and sort chromosome 6 by bivariate flow cytometry. Single chromosome 6 amplification products were confirmed with a single nucleotide polymorphism (SNP) array and verified by deep sequencing to enable assignment of both alleles at the HLA loci, defining the two haplotypes. This study exemplifies a rapid and efficient method of haplotyping that can be applied to any chromosome pair, or indeed all chromosome pairs, using a single sorting operation. The method represents a cost-effective approach to complete phasing of SNPs, which will facilitate a deeper understanding of the links between SNPs, gene regulation and protein function.

MeSH terms

  • Chromosomes, Human, Pair 6 / genetics*
  • Cytogenetic Analysis / methods*
  • Flow Cytometry / methods*
  • Genotyping Techniques / methods*
  • HLA Antigens / genetics*
  • Haplotypes*
  • Humans
  • Polymorphism, Single Nucleotide

Substances

  • HLA Antigens