人类EGFR基因突变体质控品的建立  

Establishment of human EGFR gene mutation control materials

作　　者：黄杰[1] 曲守方[1] 徐任[2] 徐超[1] 高尚先[1] 

机构地区：[1]中国食品药品检定研究院体外诊断试剂与培养基室,北京100050 [2]长春理工大学,长春130022

出　　处：《药物分析杂志》2011年第9期1758-1763,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：中国食品药品检定研究院中青年发展研究基金;项目编号2010C7

摘　　要：目的:构建了29种人类EGFR基因突变体质控品,为人类EGFR基因突变检测试剂盒的性能评价提供了可靠的质控物质。方法:根据人类EGFR全基因序列,应用Primer Premier 5.0软件在选取的序列区域设计引物,通过PCR介导的定点突变技术引入突变位点、PCR产物与克隆载体连接,转化感受态细菌,筛选阳性克隆等,建立了包含不同人类EGFR基因突变体的质控品。结果:对29种人类EGFR基因突变体质控品进行序列测定,并将测序结果进行BLAST比对,结果表明成功建立了人类EGFR基因突变体质控品。结论:本研究建立的EGFR基因突变体质控品涵盖了EGFR基因的29种突变位点,覆盖了目前最常见的EGFR基因突变类型,能够用于评价国内大多数EGFR基因突变检测试剂盒。Objective：To establish the control materials for human EGFR gene mutation including 29 different kinds for the quality control of EGFR gene mutation detection kits.Methods：According the EGFR gene sequence（including exons and introns）,designed the primers in the selected sequence region with primer primer 5.0 software.Then introduced mutations by PCR-mediated site-directed mutagenesis,ligated PCR products into the cloning vector,transformed into DH5α,and screened the positive clones by blue/white selection.Then constructed the control materials for EGFR gene mutation.Results：The control materials for human EGFR gene mutation were successfully established by sequencing the 29 recombinant plastmids.Conclusion：The control materials included 29 kinds of mutation sites,covering all the most common mutation.The control materials would satisfy the evaluation of EGFR gene mutation kits.

关 键 词：人类表皮生长因子受体(EGFR) 基因突变 质控品 PCR 

分 类 号：R917[医药卫生—药物分析学]