Development of an ARMS multiplex real-time PCR assay for the detection of HLA-B*13:01 genotype by detecting highly specific SNPs.

ABSTRACT

OBJECTIVES: HLA-B*1301 was strongly associated with Dapsone Hypersensitivity Syndrome (DHS). This study aimed to develop and validate a rapid and economical method for HLA-B*1301 genotyping. METHODS: Two tubes multiplex real-time PCR detection system comprising amplification refractory mutation system primers and TaqMan probes was established for HLA-B*1301 genotyping. Sequence-based typing was applied to validate the accuracy of the assay. RESULTS: The accuracy of the assay was 100% for HLA-B*1301 genotyping. The detection limit of the new method was 0.025 ng DNA. The positive rate of HLA-B*1301 in the Bouyei (20%, n = 50) populations was significantly higher than that in the Uighur population (4%, n = 100), Han (4.5%, n = 200), and Tibetan (1%, n = 100) ( P  < 0.05). CONCLUSION: The proposed method is rapid and reliable for HLA-B*1301 screening in a clinical setting.