甲状腺乳头状癌石蜡标本BRAF V600E基因突变检测方法的比较研究  

Comparative study of BRAF V600E gene mutation detection methods in paraffin specimens of thyroid papillary carcinoma

作　　者：易清清 梁冬雨[1] 史俊峰 沙爽 杨荣 常庆 YI Qingqing;LIANG Dongyu;SHI Junfeng;SHA Shuang;YANG Rong;CHANG Qing(Clinical Research Center,Affiliated Jiading District Central Hospital,Shanghai University of Medicine and Health Sciences,Shanghai 201800,China;Molecular Imaging Center,Shanghai University of Medicine and Health Sciences,Shanghai 201318,China;Department of Pathology,Affiliated Jiading District Central Hospital,Shanghai University of Medicine and Health Sciences,Shanghai 201800,China)

机构地区：[1]上海健康医学院附属嘉定区中心医院中心实验室,上海201800 [2]上海健康医学院分子影像中心,上海201318 [3]上海健康医学院附属嘉定区中心医院病理科,上海201800

出　　处：《国际检验医学杂志》2020年第14期1674-1677,1681,共5页International Journal of Laboratory Medicine

基　　金：上海市嘉定区农业和社会事业科研项目(JDKW-2018-W21)。

摘　　要：目的分析、比较聚合酶链反应(PCR)-荧光探针法、微滴式数字聚合酶链反应(ddPCR)法、MassARRAY核酸质谱法和Sanger测序法检测甲状腺乳头状癌患者石蜡标本中BRAF V600E基因突变阳性率。方法收集35例2013年手术切除的甲状腺乳头状癌石蜡标本(已知新鲜组织用PCR-荧光探针法检测BRAF V600E基因突变全阳性),分别采用PCR-荧光探针法、ddPCR法、MassARRAY核酸质谱法和Sanger测序法检测甲状腺乳头状癌石蜡标本中BRAF V600E基因突变情况。结果35例甲状腺乳头状癌石蜡标本PCR-荧光探针法、ddPCR法、MassARRAY核酸质谱法和Sanger测序法检测BRAF V600E基因突变阳性率分别为80.0%(28/35)、94.3%(33/35)、74.3%(26/35)和60.0%(21/35)。结论对于需要进行回顾性分析的甲状腺乳头状癌石蜡标本,ddPCR法更适用于BRAF V600E基因突变的检测。Objective To analyze and compare the positive rates of BRAF V600E gene mutation in paraffin specimens of the patients with thyroid papillary carcinoma detected by PCR-fluorescent probe method,droplet digital PCR(ddPCR)method,MassARRAY nucleic acid mass spectrometry method and Sanger sequencing method.Methods Thirty-five paraffin specimens from the patients with thyroid papillary carcinoma by surgical resection in 2013(known BRAF V600E gene mutation positive for all fresh tissues detected by PCR-fluorescent probe method)were collected.The BRAF V600E gene mutation was detected by the PCR-fluorescent probe method,ddPCR method,MassARRAY nucleic acid mass spectrometry method and Sanger sequencing method.Results The positive rates of BRAF V600E gene mutation detected by PCR-fluorescent probe method,ddPCR method,MassARRAY mass spectrometry method and Sanger sequencing method were 80.0%(28/35),94.3%(33/35),74.3%(26/35)and 60.0%(21/35)respectively.Conclusion For the paraffin specimens of thyroid papillary carcinoma needing to conduct the retrospective analysis,the ddPCR method is more suitable for the BRAF V600E gene mutation detection.

关 键 词：BRAF V600E基因突变 聚合酶链反应-荧光探针 微滴式数字聚合酶链反应 MassARRAY核酸质谱 Sanger测序 

分 类 号：R736.1[医药卫生—肿瘤]