焦磷酸测序分析表皮生长因子受体21号外显子基因突变方法的建立  

Development of pyro-sequencing test for detection of EGFR21 polymorphisms

作　　者：钟焱 周钢[2,3] 

机构地区：[1]长沙卫生职业学院,湖南长沙410100 [2]中南大学临床药理研究所,湖南长沙410078 [3]中南大学湘雅医院临床药理研究所,湖南长沙410078

出　　处：《现代预防医学》2017年第10期1862-1866,共5页Modern Preventive Medicine

摘　　要：目的建立表皮生长因子受体(Epidermal Growth Factor Receptor,EGFR)21号外显子基因突变的焦磷酸测序方法。方法用纯野生和纯突变的EGFR21质粒DNA按比例配制7份不同突变比例的DNA样本和1份正常人血液样本,对焦磷酸测序技术检测EGFR21基因突变方法进行验证,制备175例非小细胞肺癌标本g DNA,应用Qigene Pyro Mark Q24焦磷酸测序仪进行EGFR21基因的焦磷酸测序。结果建立了可同时检测EGFR基因21号外显子L858R和L861Q突变的焦磷酸测序方法,175例非小细胞肺癌标本中共检测出EGFR21突变型样本29例,总突变率为16.57%(29/175),其中L858R突变占14.29%(25/175),L861Q突变占2.28%(4/175)。结论 EGFR21基因突变的焦磷酸测序新方法具有快速、简便、灵敏度和准确度高的优点,特别适合于科研和临床批量检测的需要。Objective The aim of this study was to establish a pyro-sequencing method for detecting EGFR21 mutation L858R and L861Qin cancer tissue samples. Methods The wild and mutant EGFR21 plasmids were used to prepare seven different mutated DNA samples.And the health blood was used to prepare one normal DNA sample.The patients＇ DNA was prepared from 175 non small cell lung cancer tissue samples collected from Xiangya hospital of central south university. The target DNA sequence was amplified by PCR.And polymorphisms were detected by pyro-sequencing on Qigene PyroMark Q24. Results The pyro-sequencing method of detecting EGFR21 mutation was established. The detection rate and repetition rate were both 100%. The mutation frequencies of EGFR21 mutation in 175 non small cell lung cancer sample was 16.57%, and the positive rate of was 14.29% （25/175） for L858R mutation, 2.29% （4/19） for L861Q mutation. Conclusion The established pyro-sequencing to detect EGFR21 polymorphisms wasdemonstrated to be a rapid, simple, sensitive and accurate conventional method. And it could be performed in research and clinical application.

关 键 词：焦磷酸测序 EGFR21 非小细胞肺癌 基因突变 

分 类 号：Q33[生物学—遗传学]