焦磷酸测序法和直接测序法检测非小细胞肺癌表皮生长因子受体基因突变的比较研究

发布时间：2018-09-06 16:56

【摘要】：目的比较PCR-直接测序法和PCR-焦磷酸测序法检测表皮生成因子受体(Epidermal Growth Factor Receptor,EGFR)基因突变的一致性和差异性,探寻适应于临床检测体细胞EGFR基因突变的方法,为非小细胞癌患者的个体化用药提供理论依据。方法采用双盲的方法应用PCR-直接测序法和PCR-焦磷酸测序法检测13例(17个突变点)已知突变比例DNA样本中EGFR外显子18、19、20和21的突变情况,分析2种检测方法对各种突变的检出率和检准率;另选择实验室收集的100例非小细胞肺癌组织标本,分别采用直接测序法和焦磷酸测序法检测EGFR外显子18、19、20和21的突变情况,结果比较采用卡方检验。结果 2种检测方法对于检测EGFR18、19、20和21的结果差异有统计学差异,但焦磷酸测序法的准确度和灵敏度更高,操作更加简便。结论与直接测序法相比,PCR-焦磷酸测序法更加适用于临床检测体细胞基因突变,能够更好地识别突变比例较低的组织样本,对临床指导非小细胞肺癌TKI类药物个体化用药与预测其药物疗效都具有重要意义,值得临床进一步推广应用。
[Abstract]:Objective to compare the consistency and difference of PCR- direct sequencing and PCR- pyrosequencing in the detection of epidermal factor receptor (Epidermal Growth Factor Receptor,EGFR) gene mutations, and to explore a suitable method for clinical detection of EGFR gene mutations in somatic cells. To provide theoretical basis for individual drug use in patients with non-small cell carcinoma. Methods A double-blind method was used to detect the mutations of EGFR exon 1819 ~ (20) and EGFR exon 19 ~ (20) in 13 cases (17 mutation sites) with known mutation proportions by PCR- direct sequencing and PCR- pyrosequencing. The detection rate and accuracy of all kinds of mutations were analyzed by two methods, and the mutations in exon 1819 ~ (20) and 21 of EGFR were detected by direct sequencing and pyrosequencing, respectively, in 100 samples of non-small cell lung cancer collected in laboratory. Results the chi-square test was used. Results there were significant differences between the two methods in detecting EGFR18,19,20 and 21, but pyrosequencing was more accurate, sensitive and easy to operate. Conclusion compared with direct sequencing, PCR- pyrosequencing is more suitable for clinical detection of somatic gene mutations and can better identify tissue samples with lower mutation rate. It is of great significance to guide individual drug use of TKI and to predict its curative effect in non-small cell lung cancer (NSCLC), which is worthy of further clinical application.
【作者单位】：长沙卫生职业学院;中南大学临床药理研究所;中南大学湘雅医院临床药理研究所;
【分类号】：Q33

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