HLA-Cw allele analysis by PCR-restriction fragment length polymorphism: study of known and additional alleles

Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8803-7. doi: 10.1073/pnas.92.19.8803.

Abstract

We describe a technique for HLA-Cw genotyping by digestion of PCR-amplified genes with restriction endonucleases. Locus-specific primers selectively amplified HLA-Cw sequences from exon 2 in a single PCR that avoided coamplification of other classical and nonclassical class I genes. Amplified DNAs were digested with selected enzymes. Sixty-three homozygous cell lines from International Histocompatibility Workshop X and 113 unrelated individual cells were genotypes for HLA-Cw and compared with serology. The present protocol can distinguish 23 alleles corresponding to the known HLA-Cw sequences. Genotyping of serologically undetectable alleles (HLA-Cw Blank) and of heterozygous cells was made possible by using this method. Six additional HLA-Cw alleles were identified by unusual restriction patterns and confirmed by sequencing; this observation suggests the presence of another family of allele-sharing clusters in the HLA-B locus. This PCR-restriction endonuclease method provides a simple and convenient approach for HLA-Cw DNA typing, allowing the definition of serologically undetectable alleles, and will contribute to the evaluation of the biological role of the HLA-C locus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • B-Lymphocytes / cytology
  • Base Sequence
  • Cell Line
  • Exons / genetics
  • Gene Frequency
  • Genotype
  • HLA-C Antigens / classification
  • HLA-C Antigens / genetics*
  • Heterozygote
  • Histocompatibility Testing / methods*
  • Homozygote
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction*
  • Polymorphism, Restriction Fragment Length*
  • White People / genetics

Substances

  • HLA-C Antigens