【摘要】 目的建立一种能快速、有效检测EGFR突变的凸环引物荧光PCR新方法。方法同时用凸环引物荧光PCR技术法和Sanger测序法对混有不同比例的EGFR突变型质粒的样本进行对照检测,以对比两者灵敏度;采用两种方法对93例肺癌组织的EGFR基因进行热突变位点E746-A750del和L858R的检测,并统计其符合率。结果在93例肺癌组织的DNA中,凸环引物荧光PCR技术法和Sanger测序法检测到EGFR突变分别为29例(E746-A750del 15例,L858R14例)和28例(E746-A750del 15例,L858R 13例),符合率达96.6%,且凸环引物荧光PCR技术法能检测出混有5%突变质粒型的样本,其灵敏度达5%,比Sanger测序法高。结论凸环引物荧光PCR技术法比测序法更为简便,且灵敏度更高,易于实现,2小时内即可得出准确结果。更多还原

【Abstract】 Objective To establish a rapid and effective convex ring primer fluorescence PCR to detect EGFR gene mutation. Methods The samples mixed with different proportion of EGFR mutant plasmid were detected by convex ring primers fluorescent PCR method and Sanger sequencing method, respectively, and compare their sensitivity. Hot mutation sites E746-A750del and L858R of EGFR gene were detected in 93 cases of lung cancer tissue, and the coincidence rate were calculated. Results In 93 cases of lung cancer tissue, the convex ring primer fluorescence PCR method and Sanger sequencing method detection to EGFR mutations were 29 cases (E746-A750del 15 cases, L858R 14 cases) and 28 cases (E746-A750del 15 cases, L858R13 cases), respectively. The coincidence rate was 96.6%. The convex ring primers fluorescent PCR method could detect the samples mixed with 5% EGFR mutant plasmid, which sensitivity was 5% and higher than Sanger sequencing method. Conclusion The convex ring primers fluorescent PCR method was more sample, sensitivity and easy to realize than Sanger sequencing method, and also could get the accurate results within two hours.更多还原