对非同位素PCR-SSCP检测基因突变方法的探讨  

IMPROVEMENT OF NONISOTOPICPCRSSCP ANALYSIS TECHNIQUE IN GENE MUTATION DETECTION

作　　者：彭涛[1] 黎乐群[1] 林进令[1] 巫山[1] 陆云飞[1] 肖强[1] 曾健[1] 廖清华[1] 梁水庭[1] 

机构地区：[1]广西医科大学第一附属医院外二科

出　　处：《广西医科大学学报》1998年第1期33-36,共4页Journal of Guangxi Medical University

摘　　要：目的：建立适合本实验室条件的非同位素ＰＣＲ－ＳＳＣＰ－ＲＦＬＰ基因突变分析技术。方法：应用银染ＰＣＲ－ＳＳＣＰ技术检测石蜡包埋复发肝细胞癌（Ｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ，ＨＣＣ）组织中Ｐ５３基因第七外显子突变。采用系列步骤克服非特异扩增；分析影响ＳＳＣＰ诸参数的优化；并与ＲＦＬＰ结果相印证。结果：检测复发ＨＣＣ标本２２例，ＳＳＣＰ阳性率７７．２７％（１７／２２），２４９位点突变率７２．７２％（１６／２２），ＲＦＬＰ与ＳＳＣＰ分析结果符合率１００％；另有２例发生２４９位点以外的突变。结论：该实验流程可靠、高效。Objective:To establish nonisotopic polymerase chain reactionsingle stranded conformation polymorphism (PCRSSCP) analysis technique which is suitable for gene mutation screening of Guangxi hepatocellular carcinoma(HCC)Methods:Silver staining PCRSSCP analysis technique was used to detect P53 gene mutations in formalinfixed and paraffinembedded recurrent HCC samplesSeveral steps were chosed to obtain specific PCR products;some parameters which influence SSCP efficiency were discussed;and the result of SSCP was compared with restrictionfragmentlengthpolymorphism(RFLP) analysisResults:In all 22 cases of recurrent HHC,postive rate of SSCP analysis is 7727%(17/22),mutational rate of 249 codon is 7272%(16/22)and another 2 cases own mutations out of 249 codonConclusion:The procedure is efficient and reliable,suitable for screening of gene mutations in a great amount of samples

关 键 词：P⁵³基因 单链构象多态性分析 肝细胞癌 

分 类 号：R730.4[医药卫生—肿瘤]