| 引用本文: | 雷雅静,周丽芳,朱银焕,王安行.多重PCR-LDR法检测人体药物代谢酶基因位点多态性[J].中国现代应用药学,2021,38(4):439-444. |
| LEI Yajing,ZHOU Lifang,ZHU Yinhuan,WANG Anxing.Detection of Drug-metabolizing Enzyme Gene Locus Polymorphism in Human by Multiple PCR-LDR[J].Chin J Mod Appl Pharm(中国现代应用药学),2021,38(4):439-444. |
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| 摘要: |
| 目的 构建一套基于多重聚合酶连接反应-连接酶检测反应（polymerase chain reaction-ligase detection reaction，PCR-LDR）体系的分型法，用于同时检测多种人体主要药物代谢酶基因位点的突变。方法 选择药物代谢酶相关单核苷酸多态性（single nucleotide polymorphism，SNP）位点，设计合成各SNP位点的PCR引物和LDR探针，以人口腔黏膜细胞中提取的DNA为模板，获得PCR-LDR反应连接产物，采用ABI 3130XL进行检测分析。结果 采用本研究建立的PCR-LDR分型方法，对不同个体的14个SNP位点进行分型，分型结果与测序结果完全一致。结论 本研究建立的PCR-LDR分型方法能够同时对14个SNP位点进行一次性分型，结果准确、可靠，是一种简便有效且低成本的SNP分型方法。 |
| 关键词: 药物代谢酶 单核苷酸多态性 PCR-LDR |
| DOI：10.13748/j.cnki.issn1007-7693.2021.04.010 |
| 分类号:R969.1 |
| 基金项目: |
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| Detection of Drug-metabolizing Enzyme Gene Locus Polymorphism in Human by Multiple PCR-LDR |
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LEI Yajing, ZHOU Lifang, ZHU Yinhuan, WANG Anxing
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Hangzhou Qianyuan Enshi Gene Technology Co., Ltd., Hangzhou 310052, China
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| Abstract: |
| OBJECTIVE To develop a novel method based on polymerase chain reaction-ligase detection reaction(PCR-LDR) system for simultaneous detection of mutations in the gene locus of various major drug metabolizing enzymes in human. METHODS Single nucleotide polymorphism(SNP) sites related to drug metabolism enzymes were selected, and PCR primers and LDR probes at each SNP site were designed and synthesized. DNA extracted from human oral mucosa cells was used as template to obtain PCR-LDR reaction junction products. ABI 3130XL was used for detection and analysis. RESULTS The PCR-LDR typing method established in this study was used to classify 14 SNP sites from different individuals, and the typing results were completely consistent with the sequencing results. CONCLUSION The PCR-LDR typing method established in this study is able to perform one-time typing for 14 SNP sites at the same time with accurate and reliable results. It is a simple, effective and low-cost SNP typing method. |
| Key words: drug metabolism enzyme single nucleotide polymorphism PCR-LDR |
