以测序法为准比较HRM法、ADx-ARMS法和TaqMan探针法检测肺腺癌EGFR基因突变的敏感性  

Comparison of high resolution melting analysis,ADx-ARMS,and Taq Man PCR assays for the detection of EGFR mutations in lung adenocarcinoma tissues

作　　者：顾页[1] 黎相照[2] 薛小磊[2] 丁彦青[1] 林洁[1] 

机构地区：[1]南方医科大学基础医学院病理学系,广州510515 [2]南方医科大学南方医院病理科,广州510515

出　　处：《诊断病理学杂志》2016年第1期65-69,共5页Chinese Journal of Diagnostic Pathology

摘　　要：目的采用高分辨率熔解曲线法(HRM法)、Adx-ARMS法、Taq Man探针法和直接测序法检测肺腺癌组织中表皮生长因子受体基因(EGFR)的突变情况,并比较各方法的检测效率和敏感性。方法收集20例肺腺癌石蜡包埋肿瘤组织,其中18例为手术切除标本,2例为穿刺小标本,应用HRM法、ARMS法和Taq Man探针法分别检测EGFR基因突变,对检测结果不一致的样本采用测序法加以验证。结果 20例肺腺癌组织样本中,HRM法、ARMS法和Taq Man法分别在13例、10例和8例组织中检测到EGFR基因突变,差异不显著(P>0.05)。差异标本经测序验证后发现,HRM法可检测未知突变,而ARMS法和Taq Man探针法只能检测已知突变。对于2例穿刺小标本,HRM法和ARMS法均检测到突变,而Taq Man探针法未检测出突变,HRM法和ARMs法的检测敏感性高于Taq Man探针法。结论 HRM法、ARMS法、Taq Man探针法和测序法各有优缺点,HRM法敏感性最高,且能检测到少见突变与未知突变,因此HRM法结合测序法能更全面的检测出各种突变类型。Aims To compare the detection efficiency of HRM （high resolution melting analysis）, ADx-ARMS （scorpions amplification refractory mutation system） and Taqman probe （Taqman real time PCR） assays in EGFR mutation determination and explore a suitable clinical method for EGFR mutation detection in lung adenocarcinoma. Methods 18 cases of paraffin embedded tumor tissues and 2 case of needle biopsy sample were recruited. EGFR mutation was detected by HRM, ADx-ARMS and Taqman probe analysis in each sample. Direct sequencing was used to detect samples with inconsistent detection results of the above three different methods. Results Among the total 20 cases of lung adenoearcinoma, EGFR positive mutation was detected in 13 cases by HRM, 10 cases by ARMS, and 8 cases by Taqman PCR method. No statistical difference was found between the three methods with respect to positive mutation rate. The sequencing results shown that HRM assay can detected unknown mutations while ARMs and Taqman assays only detected known mutations. EGFR mutation was detected by HRM and ARMs method but was not detected by Taqman PCR in two needle biopsy samples. The sensibility of mutation detection of HRM and ARMs methods was higher than Taqman method. Conclusion Each method among the three assays has both strength and weakness. HRM has the highest sensibility in detecting EGFR mutation in lung adenocarcinoma and has the capability of detecting uncommon and unknown mutations. HRM assay, when combined with sequencing if necessary, is more suitable for clinical promotion and application.

关 键 词：ARMS EGFR HRM TAQMAN 测序法 非小细胞肺癌 

分 类 号：R734.2[医药卫生—肿瘤]