数字油滴PCR法与ARMS法检测结直肠癌BRAF基因突变中应用价值  

Application value of digital oil drop PCR method and ARMS method in detecting BRAF gene mutation in colorectal cancer

作　　者：姜燕平 荀延萍 傅佳仪 张仕蓉 赵妍妍 马胜林 JIANG Yanping;XUN Yanping;FU Jiayi;ZHANG Shirong;ZHAO Yanyan;MA Shenglin(Translational Medicine Research Center,Affiliated Hangzhou First People′s Hospital,Zhejiang University School of Medicine,Zhejiang Province,Hangzhou310006,China;Department of Oncology,Affiliated Hangzhou First People′s Hospital,Zhejiang University School of Medicine,Zhejiang Province,Hangzhou310006,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院转化医学研究中心,浙江杭州310006 [2]浙江大学医学院附属杭州市第一人民医院肿瘤科,浙江杭州310006

出　　处：《中国医药导报》2020年第31期21-24,39,共5页China Medical Herald

基　　金：浙江省医药卫生科技计划项目(2018KY580)。

摘　　要：目的探讨数字油滴聚合酶链式反应法(ddPCR)与扩增阻碍突变系统法(ARMS)在结直肠癌患者丝/苏氨酸特异性激酶基因(BRAF)V600E突变检测中的应用价值。方法收集2018年4月—2019年7月浙江大学医学院杭州市第一人民医院诊治的162例结直肠癌患者的手术标本,采用ARMS法和ddPCR法检测结直肠癌患者BRAF V600E突变,比较ARMS法和ddPCR法检测阳性率及一致性,分析BRAF V600E突变与结直肠癌患者临床病理特征的关系。结果ARMS法和ddPCR法检测BRAF V600E突变阳性率为8.64%(14/162)和9.26%(15/162)。对ARMS法检测BRAF V600E突变存在歧义的1例进行高通量测序,ddPCR法与高通量测序的结果一致。对应ddPCR法,ARMS法的阳性符合率、阴性符合率和一致性分别为93.33%、99.32%和99.38%。年龄越高、组织学类型低分化、临床分期Ⅲ~Ⅳ期患者BRAF V600E突变分布越高,差异有统计学意义(P<0.05)。结论ddPCR与ARMS均可用于检测结直肠癌的BRAF V600E突变,ddPCR有更高的检测效能。Objective To explore the application value of digital oil drop polymerase chain reaction(ddPCR)and amplification hindered mutation system(ARMS)in the detection of serine/threonine protein kinase(BRAF)V600E mutation in colorectal cancer patients.Methods Surgical specimens of 162 patients with colorectal cancer treated in Affiliated Hangzhou First People′s Hospital,Zhejiang University School of Medicine from April 2018 to July 2019 were collected.ARMS method and ddPCR method were used to detect BRAF V600E mutation in colorectal cancer patients,and the positive rate and consistency of ARMS and ddPCR were compared.The relationship between BRAF V600E mutation and clinicopathological characteristics of patients with colorectal cancer were analyzed.Results The positive rates of BRAF V600E mutation detected by ARMS and ddPCR were 8.64%(14/162)and 9.26%(15/162).High-throughput sequencing was performed on one case of the ambiguity of the BRAF V600E mutation detected by the ARMS method,and the result of the ddPCR method was consistent with the high-throughput sequencing result.Corresponding to the ddPCR,the positive coincidence rate,negative coincidence rate and consistency of the ARMS method were 93.33%,99.32%and 99.38%,respectively.The patients who were higher the age,the poorly differentiated histological type,in clinical stagesⅢ-Ⅳ,the higher the distribution of BRAF V600E mutations,and the differences were statistically significant(P<0.05).Conclusion Both ddPCR and ARMS can be used to detect BRAF V600E mutations in colorectal cancer,and ddPCR has higher detection efficiency.

关 键 词：结直肠癌 丝/苏氨酸特异性激酶基因V600E突变 数字油滴聚合酶链式反应法 扩增阻碍突变系统法 高通量测序 

分 类 号：R735.34[医药卫生—肿瘤]