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...
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| Veröffentlicht in: | Nature medicine 2009-11, Vol.15 (11), p.1322 |
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| creator | 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 |
| description | 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. |
| format | Article |
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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. 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| identifier | ISSN: 1078-8956 |
| ispartof | Nature medicine, 2009-11, Vol.15 (11), p.1322 |
| issn | 1078-8956 |
| language | eng |
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| source | Springer Nature - Connect here FIRST to enable access; Alma/SFX Local Collection |
| subjects | DNA sequencing Genetic aspects Health aspects Immune response Major histocompatibility complex Methods Nucleotide sequencing Physiological aspects Pyrophosphates |
| title | Major histocompatibility complex genotyping with massively paralel pyrosequencing |
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