Six Mamu-A locus alleles defined by a polymerase chain reaction sequence specific primer method

Rhesus monkeys are relevant models for human diseases and transplantation. In each case, a complete understanding of these models requires knowledge o f macaque MHC. Due to high polymorphism and multiple genes per haplotype, i r has been difficult to develop a rapid typing method for rhesus monkey MHC class I. We developed a simple and rapid PCR-SSP strategy for rhesus monke y Mamu-A locus typing. Fifty-two rhesus monkeys were included in the study. Six rhesus monkey allel-specific primer pairs were designed based on publi shed Mamu-A locus gene sequences. Allele-specific PCR products ranged in si ze from 346 to 788 bp; 5' and 3' Mamu-A locus allele specific primers were located in the second and third exons, respectively. Specific PCR product g el purification was followed by direct sequencing, without subcloning, in b oth directions. Our data showed variability in the number of Mamu-A alleles ranging from 1 to 4 per genotype. The highest frequencies were observed fo r Mamu-A*02. -A*04, and -A*03 alleles. Thus, we report here the first PCR-S SP typing method for Mamu-A*02, -03, -04, -05, -06, and -07 array of class I alleles. This technique appears to be a highly reproducible and discrimin atory method for detecting this subset of class I A locus genes in rhesus m onkeys. (C) American Society for Histocompatibility and Immunogenetics, 200 0. Published by Elsevier Science Inc.