SLA-DRB1 and -DQB1 genotyping by the PCR-SSOP-Luminex method

• Published in: Tissue antigens 2011-07, Vol.78 (1), p.49-55
• Main Authors: Ando, A., Shigenari, A., Ota, M., Sada, M., Kawata, H., Azuma, F., Kojima-Shibata, C., Nakajoh, M., Suzuki, K., Uenishi, H., Kulski, J. K., Inoko, H.
• Format: Article
• Language: English
• Subjects: Animals; Drb1 protein; Gene Frequency; Genetic Loci; Genotype; Genotyping; Haplotypes; Histocompatibility Antigens Class II - genetics; Histocompatibility Antigens Class II - immunology; Histocompatibility Testing - methods; Histocompatibility Testing - veterinary; Leukocytes; Luminex method; major histocompatibility complex; Oligonucleotide Probes - genetics; Oligonucleotides; Polymerase chain reaction; Polymerase Chain Reaction - instrumentation; Polymerase Chain Reaction - methods; polymerase chain reaction-sequence-specific oligonucleotide probe; Population genetics; Probes; Substrate Specificity - genetics; swine; Swine - genetics; Swine - immunology; swine leukocyte antigen
• ISSN: 0001-2815; 1399-0039
• DOI: 10.1111/j.1399-0039.2011.01669.x

A simple and novel genotyping method was developed to detect alleles at the swine leukocyte antigen (SLA)‐DRB1 and ‐DQB1 class II loci by using polymerase chain reaction (PCR)–fluorescently labeled sequence‐specific oligonucleotide probes (SSOPs) and Luminex 100 xMAP detection. The PCR–SSOP–Luminex method exhibited accuracy of 95% for both SLA‐DRB1 and ‐DQB1 in 6 homozygous and 16 heterozygous pig samples as confirmed by sequencing the PCR products of the same samples. In addition, 12 low‐resolution SLA class II haplotypes consisting of 7 and 9 DRB1 and DQB1 alleles were identified, respectively, in one population of 283 Landrace pigs. This genotyping method facilitates the rapid and accurate identification of two‐ or four‐digit alleles at the SLA‐DRB1 and ‐DQB1 loci.