错配碱基PCR-RFLP检测K-ras癌基因点突变  

Detection of K-ras Mutated at Codon 12 with a Modified PCR-RFLP Technique

作　　者：肖尚喜[1] 吴琳[1] 王道斌[2] 胡杰贵[1] 刘华平[1] 郑冰[1] 张军[1] 

机构地区：[1]安徽医科大学第一附属医院,合肥230022 [2]安徽医科大学病理学教研室,合肥230022

出　　处：《生物化学与生物物理进展》1997年第4期375-378,共4页Progress In Biochemistry and Biophysics

基　　金：安徽省卫生厅医学科研基金资助项目! (94 0 4)

摘　　要：错配碱基套式PCR RFLP检测K ras癌基因第 1 2位密码子点突变 ,并与一步法PCR RFLP作比较 .结果显示套式PCR RFLP可检测出 50 0细胞中的一个突变细胞 ,比一步法PCR RFLP分析的敏感性提高了 1 0 0倍 .利用该方法检测纤维支气管镜收集的标本中的突变细胞 ,结果发现 9例肺腺癌中有 5例发生了K ras癌基因第 1 2位密码子点突变 .提示该方法可行 ,值得推广应用 .An approach to detect K-ras oncogene mutated at codon 12 was devised by using a nested polymerase chain reaction strategy based on mismatched primers. A titration experiment showed that the sensitivity obtained by use of the nested PCR was 1/512. This represented the ability to detect one cell heterozygous for K-ras codon 12 mutation over 500 normal cells and was 100-fold greater than that obtained after one-step PCR. Application of this assay to 9 patients with adenocarcinoma of the lung showed 5 cases containing one or more K-ras mutation at codon 12. It is suggested that the method was practicable and worthy of application and dissemination.

关 键 词：聚合酶链反应 RAS基因 癌基因 点突变 肿瘤 

分 类 号：R730.43[医药卫生—肿瘤]