Detection of BRAF V600E mutation in fine-needle aspiration fluid of papillary thyroid carcinoma by droplet digital PCR
Detection of BRAF V600E mutation in fine-needle aspiration fluid of papillary thyroid carcinoma by droplet digital PCR
复制标题
液滴数字PCR检测甲状腺乳头状癌细针穿刺液中BRAF V600E突变
DOI：
10.1016/j.cca.2019.01.017
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发表时间：
2019-04-01
影响因子：
5
通讯作者：
Guan, Ming
中科院分区：
文献类型：
--
作者：
Xu, Xiao;Ma, Xiaowei;Guan, Ming
目的：甲状腺乳头状癌（PTC）占甲状腺癌的85%，是最常见的内分泌肿瘤。对于PTC的诊断，超声引导下细针穿刺（FNA）结合病理学评价是标准检查，BRAF V600 E突变是与PTC的发生、进展和不良临床病理特征相关的最常见分子标志物。然而，由于通过FNA获得的用于病理学评价或BRAF V600 E突变检测的肿瘤细胞量较少，因此需要更灵敏和准确的方法。方法：收集160例临床疑似甲状腺癌患者，其中146例为PTC患者，2例为滤泡性甲状腺癌（FTC），12例为甲状腺良性病变患者，均根据NCCN甲状腺癌临床实践指南进行FNA活检，并对FNA中BRAF V600 E基因突变进行检测。结果：ddPCR重复性好（CV0.01% = 14.78%，CV10% = 4.85%），检测灵敏度可达2-3拷贝/μ l（0.06%）。146例PTC患者中检测到142例BRAF V600 E突变，其中5例ARMS阴性，2例良性PTC。结论：ddPCR检测FNA液BRAF V600 E突变的敏感性和准确性均高于ARMS。
Objectives: Papillary thyroid carcinoma (PTC) accounts for 85% of thyroid carcinoma, which is the most common endocrine tumor. For the diagnosis of PTC, ultrasound-guided fine needle aspiration (FNA) with pathological evaluation is the standard test and BRAF V600E mutation is the most common molecular marker associated with the occurrence, progression and poor clinicopathological characteristics of PTC. However, because of the small amount of the tumor cells obtained by FNA for pathological evaluation or BRAF V600E mutation detection, more sensitive and accurate methods are required. Our study aimed to investigate the performance of droplet digital PCR (ddPCR) in detecting BRAF V600E mutation in FNA samples from PTC patients.Methods: One hundred and sixty suspected thyroid cancer patients were enrolled, including 146 PTC patients, 2 follicular thyroid carcinoma (FTC) and 12 benign patients, identified by FNA biopsy according to the NCCN clinical practice guidelines of Thyroid Carcinoma. ddPCR and amplification-refractory mutation system (ARMS, AmoyDx) were used to detect BRAFV600E mutation and the results were compared.Results: ddPCR had high reproducibility (CV0.01% = 14.78% and CV10% = 4.85%) and the detection sensitivity can reach 2-3 copies/mu l (0.06%). Among the 146 PTC patients, 142 BRAF V600E mutations were detected, including 5 ARMS negative patients and 2 benign cases.Conclusions: Our results demonstrated that ddPCR could be used in detecting BRAF V600E mutation from FNA fluid samples with higher sensitivity and accuracy than ARMS.