The establishment of accurate sequencing and genotyping systems of genes encoding drug metabolizing enzyme and transporter by using next generation sequencing system.

我们的目标是构建准确的药代动力学（PK）筛选系统，以揭示与药物反应性（如副作用或反应性/非反应性）相关的遗传位点。PK相关基因在超家族中具有高度同源性，在一次检测中同时扩增不同基因组区域是导致基因分型错误、序列错误和致命误诊的原因。我们将Next Generation Sequencer的对端方法与内部开发的靶向长PCR系统相结合，鉴定了28个编码药物代谢酶和药物转运体的基因CYP1A2、CYP2A6、CYP2C8、CYP2C18、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4、OAT1、OAT2、OAT3、OCT1、OCT2、OCTN1、OCTN2、OATP1、OATP2、MATE1、MATE2、MRP1、MRP2、MDR1、PEPT1、PEPT2、PMAT、TPMT和UGT1A1在不同区域的高度同源性。此外，我们还构建了一个基于Invader法和长PCR相结合的常规基因分型系统，该系统几乎具有短变异（94%）。我们计划与其他专注于药物反应的研究人员合作进行常规筛查。

Our aim is to construct accurate screening system for pharmacokinetics(PK) to uncover genetic loci associate with drug responsiveness such as side effects or responder/non-responder. Genes related to PK well known to recognize high homology within superfamily, simultaneous amplification of different genomic regions in one test is causes of incorrect genotyped, incorrect sequence, and a fatal mistake in diagnosis. We identified all variations in 28 genes encoding drug metabolizing enzymes and drug transporters, CYP1A2, CYP2A6, CYP2C8, CYP2C18, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, OAT1, OAT2, OAT3, OCT1, OCT2, OCTN1, OCTN2, OATP1, OATP2, MATE1, MATE2, MRP1, MRP2, MDR1, PEPT1, PEPT2, PMAT, TPMT, and UGT1A1, recognized high homology in different regions, based on the combination the pair-end method of Next Generation Sequencer with an internally developed target specific long PCR system. In addition, we constructed a routine genotyping system of almost short variations(94%) based on combination Invader method and the long PCR. We planning to perform the routine screening in collaboration with other researchers focus on drug responsiveness.