热裂解法在微量甲状腺细针穿刺样本行分子检测中的应用  

Application of two nucleic acid extraction methods in BRAF V600E detection of thyroid fine needle aspiration samples

作　　者：潘苡晴 吕纬苓 方庆全[1] 郑捷[1] Pan Yiqing;Lyu Weiling;Fang Qingquan;Zheng Jie(Department of Pathology,the First Affiliated Hospital of Xiamen University,Xiamen 361003,China;College of Integrated Traditional Chinese and Western Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 351001,China)

机构地区：[1]厦门大学附属第一医院病理科,厦门361003 [2]福建中医药大学中西医结合学院,福州351001

出　　处：《实用医技杂志》2025年第4期275-279,共5页Journal of Practical Medical Techniques

摘　　要：目的探讨热裂解法在提取甲状腺微小结节的细针穿刺样本液基细胞学制片后剩余微量样本的DNA进行BRAF V600E突变检测方面的应用价值。方法收集2023年1-6月厦门大学附属第一医院甲状腺微小结节细针穿刺样本进行液基细胞学制片后的剩余标本60份,随机分为2组,每组30例,分别采用热裂解法与离心柱法提取DNA,比较2种提取方法的产物DNA浓度、纯度、循环阈值(CT)和PCR扩增结果有效率、BRAF V600E检测突变率;以术后石蜡包埋组织的BRAF V600E检测结果为金标准,比较2种提取方法下甲状腺穿刺液BRAF V600E检测结果的符合率。结果热裂解法提取DNA产物的浓度高于离心柱法[25.20(21.58,61.73)ng/μl比4.85(3.60,12.83)ng/μl,P<0.001],纯度低于离心柱法[0.92(0.85,1.03)比1.71(1.43,1.86),P<0.001];2组提取产物进行BRAF V600E检测,热裂解法扩增结果有效率显著高于离心柱法[100.0%(30/30)比73.3%(22/30),P=0.008];2种提取方法下BRAF V600E检测突变率差异无统计学意义(60.0%和60.0%,P=1.000);对照术后病理组织的BRAF V600E检测结果,热裂解法下甲状腺穿刺样本BRAF V600E检测结果的符合率高于离心柱法[93.3%(28/30)比73.3%(22/30),P=0.038]。结论采用热裂解法提取甲状腺细针穿刺样本液基细胞学制片后剩余微量样本的DNA进行BRAF V600E检测,成本较低廉,操作较简单,流程用时较短,所提取的DNA浓度较高,PCR检测有效率较高,假阴性较低,适用于甲状腺微小结节术前细针穿刺液同时行液基细胞学检查和分子检测。Objective To investigate the application value of thermal lysis method in extracting DNA from trace residual samples of thyroid fine-needle aspiration specimens after liquid-based cytological preparation,specifically for BRAF V600E mutation detection.Methods Six residual samples from thyroid micronodule fine-needle aspiration were collected after liquid-based cytology smears were prepared,and were randomly divided into two groups with 30 cases each.DNA was extracted using the thermal lysis method and the centrifugal column method respectively.The concentration and purity of the extracted DNA,the CT value and effective rate of PCR amplification,and the BRAF V600E mutation detection rate were compared.Taking the BRAF V600E mutation detection results of postoperative paraffin-embedded tissues as the gold standard,we compared the concordance rates of BRAF V600E detection results under the two extraction methods.Results The concentration of DNA extracted by the thermal lysis method was higher than that by the centrifugal column method[25.20(21.58,61.73)ng/μl vs.4.85(3.60,12.83)ng/μl,P<0.001],whereas the purity was lower[0.92(0.85,1.03)vs.1.71(1.43,1.86),P<0.001].The effective rate of PCR amplification for BRAF V600E detection by thermal lysis method was significantly higher than that by centrifugal column method[100.0%(30/30)vs.73.3%(22/30),P=0.008].There was no difference in the BRAF V600E mutation detection rate between the two methods(60.0%vs.60.0%,P=1.000).The concordance rate of BRAF V600E detection in fine-needle aspiration samples by thermal lysis method was higher than that by centrifugal column method when compared with postoperative pathological tissue BRAF V600E detection results[93.3%(28/30)vs.73.3%(22/30),P=0.038].Conclusion The thermal lysis method for extracting DNA from trace residual samples of thyroid fine-needle aspiration after liquid-based cytology preparation for BRAF V600E detection is cheaper,simpler,and faster,yielding higher DNA concentration,higher PCR detection efficiency,and lower false

关 键 词：甲状腺 活组织检查 细针 甲状腺癌 乳头状 热裂解法 离心柱法 

分 类 号：R736.1[医药卫生—肿瘤]