分子诊断与治疗杂志
分子診斷與治療雜誌
분자진단여치료잡지
JOURNAL OF MOLECULAR DIAGNOSIS AND THERAPY
2013年
2期
86-90
,共5页
EGFR%凸环引物荧光 PCR 方法%Sanger测序法
EGFR%凸環引物熒光 PCR 方法%Sanger測序法
EGFR%철배인물형광 PCR 방법%Sanger측서법
EGFR%Convex ring primers fluorescent PCR method%Sanger sequencing method
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,L858R 14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.
目的建立一種能快速、有效檢測 EGFR 突變的凸環引物熒光 PCR 新方法.方法同時用凸環引物熒光 PCR 技術法和 Sanger 測序法對混有不同比例的 EGFR 突變型質粒的樣本進行對照檢測,以對比兩者靈敏度;採用兩種方法對93例肺癌組織的 EGFR 基因進行熱突變位點 E746-A750del 和 L858R的檢測,併統計其符閤率.結果在93例肺癌組織的 DNA 中,凸環引物熒光 PCR 技術法和 Sanger 測序法檢測到 EGFR 突變分彆為29例(E746-A750del 15例,L858R 14例)和28例(E746-A750del 15例, L858R 13例),符閤率達96.6%,且凸環引物熒光 PCR 技術法能檢測齣混有5%突變質粒型的樣本,其靈敏度達5%,比 Sanger 測序法高.結論凸環引物熒光 PCR 技術法比測序法更為簡便,且靈敏度更高,易于實現,2小時內即可得齣準確結果.
목적건립일충능쾌속、유효검측 EGFR 돌변적철배인물형광 PCR 신방법.방법동시용철배인물형광 PCR 기술법화 Sanger 측서법대혼유불동비례적 EGFR 돌변형질립적양본진행대조검측,이대비량자령민도;채용량충방법대93례폐암조직적 EGFR 기인진행열돌변위점 E746-A750del 화 L858R적검측,병통계기부합솔.결과재93례폐암조직적 DNA 중,철배인물형광 PCR 기술법화 Sanger 측서법검측도 EGFR 돌변분별위29례(E746-A750del 15례,L858R 14례)화28례(E746-A750del 15례, L858R 13례),부합솔체96.6%,차철배인물형광 PCR 기술법능검측출혼유5%돌변질립형적양본,기령민도체5%,비 Sanger 측서법고.결론철배인물형광 PCR 기술법비측서법경위간편,차령민도경고,역우실현,2소시내즉가득출준학결과.
@@@@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.
