Major histocompatibility complex genotyping with massively parallel pyrosequencing

Roger W. Wiseman, Julie A. Karl, Benjamin N. Bimber, Claire E. O'Leary, Simon M. Lank, Jennifer J. Tuscher, Ann M. Detmer, Pascal Bouffard, Natalya Levenkova, Cynthia L. Turcotte, Edward Szekeres, Chris Wright, Timothy Harkins, David H. O'Connor

Research output: Contribution to journalArticlepeer-review

121 Scopus citations


Major histocompatibility complex (MHC) genetics dictate adaptive cellular immune responses, making robust MHC genotyping methods essential for studies of infectious disease, vaccine development and transplantation. Nonhuman primates provide essential preclinical models for these areas of biomedical research. Unfortunately, given the unparalleled complexity of macaque MHCs, existing methodologies are inadequate for MHC typing of these key model animals. Here we use pyrosequencing of complementary DNA-PCR amplicons as a general approach to determine comprehensive MHC class I genotypes in nonhuman primates. More than 500 unique MHC class I sequences were resolved by sequence-based typing of rhesus, cynomolgus and pig-tailed macaques, nearly half of which have not been reported previously. The remarkable sensitivity of this approach in macaques demonstrates that pyrosequencing is viable for ultra-high-throughput MHC genotyping of primates, including humans.

Original languageEnglish (US)
Pages (from-to)1322-1326
Number of pages5
JournalNature medicine
Issue number11
StatePublished - Nov 2009
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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