双环探针特异引物荧光聚合酶链反应法检测非小细胞肺癌表皮生长因子受体基因突变

目的探讨双环探针特异引物荧光聚合酶链反应法(BPSP-qPCR)检测非小细胞肺癌( NSCLC)表皮生长因子受体(EGFR)基因突变的敏感性和稳定性。方法采用BPSP-qPCR法检测NSCLC中EGFR基因第18～21号外显子突变,分析突变与临床病理特征、不同检测样本间的关系。结果265例NSCLC样本中19-del和（或）L858R突变占30.2％( 80/265)。资料完整的184例中,女性、不吸烟者、腺癌有更高的19-del和（或）L858R突变率,分别为39.7％ (31/78)、41.0％ (43/105)、37.8％ (51/135) (P ＜0.05);T790M合并19-del和（或）L858R占3.3％(6/184);男性转移灶、女性和不吸烟者胸腔积液样本有较高的19-del和（或）L858R阳性突变率,分别为29.6％( 8/27)、42.9％(9/21)和40.7％(11/27),而非腺癌转移灶为0(P＞0.05)。结论BPSP-qPCR法检测EGFR基因突变稳定、敏感。EGFR基因敏感突变多见于女性、非吸烟、腺癌,胸腔积液样本和转移灶少量样本能够检出EGFR突变,指导临床治疗。

Detection of epidermal growth factor receptor gene mutations in non-small cell lung cancer using bi-loop probe specific primer quantitative PCR

Objective To investigate the sensitivity of bi-loop probe and specific primer quantitative PCR (BPSP-qPCR) in the detection of epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC). Methods BPSP-qPCR was employed to examine the presence of mutations of EFGR exon 19 through 21. Correlation of the mutations with clinicopathological characteristics and types of tumor samples were performed. Results In the cohort of 265 specimens, 30.2％ ( 80/265 )mutations were found to be 19-del and/or L858R. Females (39.7％, 31/78 ), non-smokers (41.0％,43/105) and adenocarcinoma patients (37.8％ ,51/135) had a higher mutation rate (P＜0.05) among 184 patients whose profiles were available. T790M combined with 19-del and/or L858R accounted for 3.3％ (6/184) of the mutations. Male metastatic tumors (29.6％ ,8/27), pleural fluids of females (42.9％,9/21 ) and non-smokers (40.7％, 11/27) were found to have higher percentage of 19-del and/or L858R mutations, in contrast, no mutations were found in the metastatic lesions of non-adenocarcinoma patients (P ＞0.05). Conclusions BPSP-qPCR is a robust method in detection of EGFR mutations with high consistency and sensitivity. The difference of EGFR mutations in primary tumors, metastatic lesions and pleural fluids suggests that EGFR tyrosine kinase inhibitors (EGFR-TKI) treatment may have variable treatment effects depending on the tumor sites.