【摘要】 目的探讨ARMS法和Sanger测序法检测KRAS基因突变的差异。方法整理收集南方医院2013—2016年间手术切除的113例结肠癌标本,同时应用ARMS法和Sanger测序法检测KRAS基因突变,分析两种方法的检测结果以及检出模式,考察两种方法的检测差异程度。结果 113例结肠癌手术标本ARMS法和Sanger测序法检出KRAS基因突变的阳性率分别为47.8%(54/113)和34.5%(39/113),两种方法检测结果阳性符合率为72.2%(39/54)。ARMS法检测总突变率显著高于Sanger测序法(P<0.05);ARMS法检测4种单碱基的突变率均高于Sanger测序法(P>0.05)。其中4例Sanger测序法检测结果为(+),ARMS法为(-);19例标本ARMS法检测结果为(+),用Sanger测序法检测结果为(-);另有1例标本用ARMS法检测到有GGT>GAT(G12D)和GGC>GAC(G13D)双突变。结论 ARMS法和Sanger测序法各有优缺点,2种方法检测,其结果一致。ARMS法能检测出的阳性病例更多,且结果差异显著,而Sanger测序法检测突变种类较多。综合考虑临床检测结肠癌手术标本KRAS基因突变应以ARMS法为主,Sanger测序法作为补充,可提高KRAS基因突变检出率。更多还原

【Abstract】 Objective To compare the difference of KRAS gene mutation between ARMS method and Sanger sequencing.Methods 113 colon cancers were enrolled in this study from the Southern Hospital during 2013-2016. ARMS method and Sanger sequencing were performed to detect KRAS gene mutations. The difference in frequency of gene mutation between the two methods was statistically analyzed by SPSS 20 statistics software. Results KRAS gene mutation was detected in 113 cases of colon cancer specimens by ARMS method and Sanger sequencing. The positive rates were 47. 8( 54/113) and 34. 5%( 39/113) respectively,resulting in the total coincidence rate was 72. 2%( 39/54).Based on the statistical results,we come to the conclusion that the total mutation rate of ARMS method was significantly higher than that of Sanger sequencing( P < 0. 05); The mutation rate of 4 kinds of single bases detected by ARMS method was higher than that of Sanger sequencing( P > 0. 05). 4 positive samples with Sanger sequencing were negative with ARMS-PCR; ARMS method test results were positive in 19 cases,and the result of Sanger sequencing test was negative.In another sample,double mutations were detected in GLY12 ASP( GGT > GAT) and GLY13 ASP( GGC > GAC) by ARMS method. Conclusion Each of the two methods has its advantages and disadvantages. The results are consistent with 2 methods of detection. Considering the clinical examination,the KRAS gene mutation in colon cancer specimens should be based on ARMS method,and Sanger sequencing method is used as a supplement to improve the detection rate of KRAS gene mutation.更多还原