Comparison of ARMS method with Sanger sequencing for detection of KRAS mutation in colon cancer

XUE Xiao-lei; ZHAO Liang; PAN Zhi-hua; LI Xiang-zhao; DING Yan-qing

doi:10.3969/j.issn.1007-8096.2018.02.005

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 GLY12ASP(GGT ＞ GAT) and GLY13ASP (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.