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)筛查系统，以揭示与药物反应相关的遗传位点，如副作用或应答者/无应答者。众所周知，与PK相关的基因识别超家族内的高度同源性，在一次测试中同时扩增不同基因组区域是导致错误基因分型、错误序列和致命诊断错误的原因。我们鉴定了编码药物代谢酶和药物转运蛋白的28个基因的所有变异，根据下一代测序器的对端方法和内部开发的靶标长片段聚合酶链式反应系统，在不同的区域识别出高度同源性的基因：CYP1A2、CYP2A6、CYP2C8、CYP2C18、CYP2C9、CYP2C19、CYP2D6、CYP2E1、CYP3A4、OAT1、OAT2、OAT3、OCT1、OCT2、OCTN1、OCTN2、OATP1、Mate2、MRP1、MRP2、MDR1、PEPT1、PEPT2、pmAt、TPMT和UGT1A1。此外，我们还构建了一个几乎短变异(94%)的常规基因分型系统，该系统基于组合入侵者方法和长PCR。我们计划与其他研究人员合作进行常规筛查，重点放在药物反应上。

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.