Development of a rapid and reliable single-tube multiplex real-time PCR method for HLA-A*24:02 genotyping.

Aim:HLA-A*24:02 is significantly associated with cutaneous adverse drug reactions caused by aromatic antiepileptic drugs. Here, we aimed to establish a fast and reliable detection method for HLA-A*24:02 genotyping. Methods: A single-tube multiplex quantitative real-time polymerase chain reaction(qPCR) assay for HLA-A*24:02 genotyping was established by combining allele-specific primers with TaqMan probes. Results: A 100% concordance was observed between qPCR and SBT result in 106 Han subjects. The detection limit of the new method was 0.05ng DNA. The positive rate of HLA-A*24:02 in Tibetans (55.6%, n=81) was significantly higher than those in Han (34%, n=106), Uighur (27.5%, n=102), Bouyei (25.9%, n=116) and Miao populations (26.5%, n=113). Conclusion: The newly established qPCR assay was reliable for HLA-A*24:02 screening in clinical applications.