Analysis of primate major histocompatibility complex (MHC)-DQA1 locus by PCR-single strand conformation polymorphism (SSCP).
Major histocompatibility complex (Mhc) gene products play an important role in the immune responses against pathogens and autoimmunity, disease resistance and transplantation. Non-human primates (NHPs) are increasingly being utilized as models to test the safety and efficacy of candidate vaccines. Mhc typing of NHPs is an important component of the vaccine trial studies and in the investigations of any associations between Mhc alleles and disease. Routine typing of primate Mhc alleles has been hampered by unavailability of well characterised immunological reagents. In this study, we have used PCR amplification and SSCP for screening polymorphisms in the primate DQA1 locus. Using this technique, 9 African primate species (36 individuals) were analyzed. Ten individuals showed three or four electrophoretic band patterns and the rest two-band patterns indicating this technique can be used to discriminate homozygous and heterozygous individuals prior to DNA sequencing. This method may also be used to screen primates for Mhc-DQA1 allelic polymorphism. However, practical application of this technique for routine typing of primate Mhc-DQA1 alleles depends on availability of adequate nucleotide sequence information.
Mwenda
J M
JM
Institute of Primate Research, National Museums of Kenya, Karen Nairobi, Kenya.
Hashiba
K
K
Bambra
C S
CS
Shotake
T
T
D000483:Alleles; D000818:Animals; D006224:Cricetinae; D006683:HLA-DQ Antigens / Q000235:genetics*; D059848:HLA-DQ alpha-Chains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011323:Primates / Q000235:genetics* / Q000276:immunology
D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen