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

Zohreh Tatari Calderone, Catherine Fortier, Vlasta Bobrynina, Pascale Loiseau, Dominique Charron, Colette Raffoux

Research output: Contribution to journalArticle

27 Citations (Scopus)

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 genotyped 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.

Original languageEnglish
Pages (from-to)8803-8807
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number19
DOIs
Publication statusPublished - 12 Sep 1995
Externally publishedYes

Fingerprint

Restriction Fragment Length Polymorphisms
Alleles
Polymerase Chain Reaction
DNA Restriction Enzymes
HLA-C Antigens
MHC Class I Genes
Histocompatibility
HLA-B Antigens
DNA Fingerprinting
Serology
Digestion
Exons
Education
Cell Line
DNA
Enzymes
Genes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

HLA-CW allele analysis by PCR-restriction fragment length polymorphism : Study of known and additional alleles. / Tatari Calderone, Zohreh; Fortier, Catherine; Bobrynina, Vlasta; Loiseau, Pascale; Charron, Dominique; Raffoux, Colette.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 19, 12.09.1995, p. 8803-8807.

Research output: Contribution to journalArticle

Tatari Calderone, Zohreh ; Fortier, Catherine ; Bobrynina, Vlasta ; Loiseau, Pascale ; Charron, Dominique ; Raffoux, Colette. / HLA-CW allele analysis by PCR-restriction fragment length polymorphism : Study of known and additional alleles. In: Proceedings of the National Academy of Sciences of the United States of America. 1995 ; Vol. 92, No. 19. pp. 8803-8807.
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