Study of MICA alleles in 201 African Americans by multiplexed single nucleotide extension (MSNE) typing

Yanzheng Zhang, Mei Han, Robert Vorhaben, Chris Giang, Bhavna Lavingia, Peter Stastny

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

We have developed a method for major histocompatibility complex class I chain-related gene A (MICA) genotyping using multiplexed single nucleotide extension (MSNE) and flow cytometric analysis of an array of fluorescent microspheres. This technique employs a polymerase chain reaction-derived target DNA containing all the polymorphic sites of MICA, synthetic complementary primers, biotinylated dideoxynucleotide triphosphate, fluorescent reporter molecules (streptavidinphycoerythrin), and thermophilic DNA polymerase. Genomic DNA was amplified by MICA locus-specific primers and the MSNE reactions were carried out in the presence of 30 MSNE primers used to assay polymorphisms in exons 2, 3, and 4 of the MICA genes. Thirty-two previously typed cell lines were used as reference material. The MICA gene frequencies among 201 African-American unrelated donors were determined. Of 51 previously known alleles, 18 were observed in AfricanAmericans, compared to 16 that were found in North American Caucasians and 9 in South American Indians, suggesting a more diversified allelic distribution in African-Americans. MICA*00201 and MICA*00801 were the two most frequent alleles in African-Americans. We observed a high degree of linkage disequilibrium between certain alleles of MICA and of human leukocyte antigen-B in the African-American population. The methodology described here offers a powerful new approach to DNA typing of the MICA alleles. Human Immunology 64,

Original languageEnglish (US)
Pages (from-to)130-136
Number of pages7
JournalHuman Immunology
Volume64
Issue number1
DOIs
StatePublished - Jan 1 2003

Keywords

  • African Americans
  • Allelic diversity
  • Linkage disequilibrium
  • MICA
  • MSNE

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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