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Major histocompatibility complex genotyping with massively parallel pyrosequencing.

Nat. Med.2009 Nov;15(11):1322-6. Epub 2009 Oct 11
Roger W Wiseman 1 , 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
Roger W Wiseman 1 , 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
+ et al

[No authors listed]

Author information
  • 1 Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, Wisconsin, USA.

摘要


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.

原始数据


 保存测序数据
Sample name
Organism Experiment title Sample type Library instrument Attributes
origin
Rhesus macaque MHC class I cDNA sequences
Macaca mulatta Indian origin rhesus macaque MHC class-I amplicons AMPLICON 454 GS FLX India