PS-TaqMan PCR检测巯基嘌呤甲基转移酶基因多态性

目的为临床开展巯基嘌呤类药物的个体化用药需要,发展一种适合临床实验室检测TPMT的基因多态性的方法。方法改进TaqMan错配扩增突变检测法（TaqMAMA）,建立引物特异性TaqMan聚合酶链反应（PS-TaqManPCR）,检测245例临床样本的TPMT＊3C突变,结果用双向测序方法验证。结果 PS-TaqManPCR能够准确检测TPMT＊3C突变,TMPT＊1/＊3C基因型频率是2.4%。结论成功发展PS-TaqManPCR作为临床检测TPMT基因多态性的方法 ,该方法可靠、高通量且效率高,适合临床实验室对样本大规模检测的需要。

Developing a novel SNP detection method,PS-TaqMan PCR,to detect TPMT genetype in Chinese

Objective In vitro studies indicate that TPMT variant has an impaired capacity for mercaptopurine drug metabolism.Therefore,an efficient detection for this mutation may be important for clinical dose adjustment.To develop an appropriate tool for TMPT polymorphism detection in clinical laboratory.Methods The previous TaqMan mismatch amplification mutation assay（TaqMAMA） was modified to primer special-TaqMan polymerase chain reaction（PS-TaqMan PCR） to satisfy the high-through requirements.254 genomic DNAs of south Chinese were detected TPMT＊3C to test the detection system.The results were checked by bi-directional sequencing.Results PS-TaqMan PCR could exactly genotype genomic template.Among the 254 genomic DNAs,6 heterozygotes and 248 wild-type homozygotes were detected and confirmed by bi-directional sequencing.Conclusion PS-TaqMan PCR was successfully developed for TPMT＊3C detection.The reliable,high-throughput and efficient tool could satisfy the requirements of clinical laboratory.