Rapid high-resolution MHC class I genotyping of Chinese rhesus macaques by capillary reference strand-mediated...

Area of Science:

• Immunology
• Genetics
• Primate Models

Background:

• Rhesus macaques are crucial models for human disease research and vaccine development.
• Individual differences in disease susceptibility in macaques are linked to major histocompatability complex (MHC) class I sequences.
• A need exists for rapid, cost-effective, high-resolution MHC genotyping in macaques.

Purpose of the Study:

• To adapt and validate a capillary electrophoresis-based method for high-resolution MHC class I genotyping in rhesus macaques.
• To assess the concordance of the new method with established sequencing techniques.
• To determine the frequency of specific MHC class I sequences in a cohort of Chinese rhesus macaques.

Main Methods:

• Adaptation of reference strand-mediated conformational analysis (RSCA) for macaque MHC class I genotyping using an ABI 3130xl genetic analyzer.
• Validation of RSCA by comparing results with complementary DNA cloning and sequencing data for 14 MHC class I sequences in 12 macaques.
• Genotyping of an additional 25 Chinese rhesus macaques to determine MHC sequence frequencies.

Main Results:

• RSCA demonstrated over 98% concordance with cloning and sequencing methods for MHC class I genotyping.
• The method confirmed MHC haplotype sharing between macaques, consistent with prior microsatellite analysis.
• Frequencies of 14 MHC class I sequences in 25 macaques ranged from 5% to 32%, with Mamu-A1*2601 being the most prevalent.

Conclusions:

• Capillary RSCA is a rapid, cost-effective, and accurate method for high-resolution MHC class I genotyping in rhesus macaques.
• This technique facilitates the evaluation of MHC class I genotypes and their association with disease susceptibility.
• The findings provide a valuable tool for advancing research in primate immunology and infectious diseases.