Major histocompatibility complex genotyping with massively paralel pyrosequencing

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...

Full description

Saved in:
Bibliographic Details
Published inNature medicine Vol. 15; no. 11; pp. 1322 - 1327
Main Authors Wiseman, Roger W, Karl, Julie A, Bimber, Benjamin N, O'Leary, Claire E, Lank, Simon M, Tuscher, Jennifer J, Detmer, Ann M, Bouffard, Pascal, Levenkova, Natalya, Turcotte, Cynthia L, Szekeres, Jr., Edward, Wright, Chris, Harkins, Timothy, O'Connor, David H
Format Journal Article
LanguageEnglish
Published Nature Publishing Group 01.11.2009
Subjects
DNA sequencing
Genetic aspects
Health aspects
Immune response
Major histocompatibility complex
Methods
Nucleotide sequencing
Physiological aspects
Pyrophosphates
United States
Online AccessGet full text

Cover

More Information
Summary: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 pigtailed 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.
ISSN:1078-8956
1546-170X