基于实时荧光定量PCR的ARMS法检测BRAFV600E点突变在甲状腺微小乳头状癌中的早期诊断价值  

Value of ARMS based on real-time fluorescent quantitative PCR detecting BRAF V600E mutation in the early diagnosis of papillary thyroid microcarcinoma

作　　者：丁金旺[1] 罗定存[1] 荀延萍 徐益平[2] 王炜[1] 叶柳青[3] 

机构地区：[1]杭州市第一人民医院南京医科大学附属杭州医院,浙江杭州310006 [2]杭州市肿瘤医院,浙江杭州310002 [3]浙江省肿瘤医院,浙江杭州310022

出　　处：《中国卫生检验杂志》2016年第13期1888-1891,共4页Chinese Journal of Health Laboratory Technology

基　　金：杭州市重大科技创新专项项目(20131813A08);杭州市医药卫生科技计划项目(2012B004;2013Z04);杭州市科技计划项目(20160533B05)

摘　　要：目的探讨BRAF V600E点突变在甲状腺微小乳头状癌(PTMC)和甲状腺良性病变中的表达状况,并评估其在PTMC中的早期诊断价值。方法采用基于实时荧光定量PCR技术的扩增阻滞突变系统(ARMS)法检测并比较104例PTMC和42例甲状腺良性病变中BRAF V600E点突变的表达差异,分析BRAF V600E点突变与PTMC临床病理特征的相关性,计算BRAF V600E点突变对鉴别PTMC及甲状腺良性病变的各项临床诊断指标。结果 104例PTMC的BRAF V600E点突变率为83.65%,而42例甲状腺良性病变的突变率仅为2.38%,BRAF V600E点突变率在2组间差异有统计学意义(χ2=82.531,P<0.05)。BRAF V600E点突变与PTMC患者的性别、年龄、肿瘤大小、病灶数目、双侧累及、被膜侵犯、淋巴结转移及TNM分期均无相关性(P>0.05)。ARMS法检测BRAF V600E点突变对诊断PTMC的敏感度为83.65%,特异度为97.62%,准确度为87.67%,约登指数为0.812 7,阳性预测值为98.86%,阴性预测值为70.69%。结论 BRAF V600E点突变是诊断PTMC的重要标记物,基于实时荧光定量PCR的ARMS法检测BRAF V600E点突变对PTMC具有良好的早期诊断价值。Objective To explore the expression of BRAF V600 E mutation in the papillary thyroid microcarcinoma（ PTMC） and thyroid benign lesions,and to evaluate its early diagnostic value in PTMC. Methods The method of ARMS based on real-time fluorescent quantitative PCR was adopted to detect and compare the difference of BRAF V600 E mutation between in 104 PTMCs and 42 cases of thyroid benign lesions. Correlation between BRAF V600 E mutation and clinical pathological features of PTMC was also analyzed. Finally,various diagnostic indicators of BRAF V600 E mutation was calculated to identify PTMC and thyroid benign lesions. Results The mutation rate of BRAF V600 E in 104 PTMC was 83. 65%,whereas that of 42 thyroidbenign lesions was only 2. 38%,and the mutation rate differences of BRAF V600 E between the two groups was statistically significant（ χ2= 82. 531,P〈0. 05）. There was no association of the BRAF V600 E mutation with the PTMC patient＇s gender,age,tumor size,number of lesions,bilateral involvement,membrane invasion,lymph node metastasis and TNM stage（ P〉0. 05）. The diagnostic sensitivity of ARMS method detecting BRAF V600 E mutation for PTMC was 83. 65%,specificity was 97. 62%,accuracy was 87. 67%,Youden index was 0. 812 7,positive predictive value was 98. 86%,and negative predictive value was70. 69%. Conclusion BRAF V600 E mutation is one of the important markers for the diagnosis of PTMC,and detection of BRAF V600 E mutation by PCR-ARMS shows significant value for the early diagnosis of PTMC.

关 键 词：甲状腺微小乳头状癌 BRAF V600E点突变 扩增阻滞突变系统 标记物 

分 类 号：R736.1[医药卫生—肿瘤]