Development of a Rapid and Reliable Single-Tube Multiplex Real-Time PCR Method for HLA-A24:02 Genotyping

• Published in: Pharmacogenomics Vol. 20; no. 11; pp. 803 - 812
• Main Authors: Wang, Yanxia, Zhang, Tingting, Zhang, Lirong, Pei, Yanrui, Zhao, Lili, Li, Yanwei, Liu, Lin, Wang, Huijuan
• Format: Journal Article
• Language: English
• Published: England Future Medicine Ltd 01.07.2019
• Subjects: Alleles; Anticonvulsants - adverse effects; Anticonvulsants - therapeutic use; Antiepileptic agents; Asian Continental Ancestry Group; Carbamazepine - adverse effects; Carbamazepine - therapeutic use; Deoxyribonucleic acid; DNA; DNA probes; Epilepsy - complications; Epilepsy - drug therapy; Epilepsy - genetics; Female; Genetic Predisposition to Disease; Genotype; Genotyping; Genotyping Techniques - methods; Healthy Volunteers; Histocompatibility antigen HLA; HLA-A24 Antigen - genetics; Humans; Male; Methods; Multiplex Polymerase Chain Reaction - methods; Mutation; Polymerase chain reaction; Population; Primers; Real-Time Polymerase Chain Reaction - methods; Skin Diseases - chemically induced; Skin Diseases - genetics; Skin Diseases - pathology; Systematic review; Therapeutic applications; China; ARMS; aromatic antiepileptic drugs (AEDs); qPCR; cutaneous adverse drug reactions (cADRs)
• ISSN: 1462-2416; 1744-8042; 1744-8042
• DOI: 10.2217/pgs-2019-0052

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 genotyping. A single-tube multiplex quantitative real-time polymerase chain reaction (qPCR) assay for genotyping was established by combining allele-specific primers with TaqMan probes. A 100% concordance was observed between qPCR and SBT result in 106 Han subjects. The detection limit of the new method was 0.05 ng DNA. The positive rate of 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). The newly established qPCR assay was reliable for screening in clinical applications.