实时荧光定量PCR和测序法检测非小细胞肺癌EGFR基因突变的比较

EGFR Mutations Detection in Non-small Cell Lung Cancer Tissues by Real-time PCR and DNA Sequencing

背景与目的以厄洛替尼(Erlotinib)为代表的小分子表皮生长因子受体酪氨酸激酶抑制剂在治疗非小细胞肺癌已经取得很好的疗效。许多研究发现表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变与酪氨酸激酶抑制剂的疗效密切相关。本研究目的是通过比较荧光定量PCR(Real-time PCR)和DNA测序法检测EGFR基因突变的一致性,建立适用于临床的EGFR基因突变检测方法。方法应用荧光定量PCR(Real-time PCR)及DNA测序法检测103例非小细胞肺癌标本的EGFR外显子19和21的基因突变,结果比较采用卡方检验。结果两种方法对EGFR外显子19和21的基因突变的检测结果无统计学差异,且荧光定量PCR法比测序法更快捷和灵敏。结论与测序法相比,荧光定量PCR更适用于临床检测。

Background and objective Small molecule tyrosine kinase inhibitors （TKIs）,such as gefitinib and erlotinib that target the kinase domain of epidermal growth factor receptor （EGFR） are making successful progression for advanced non-small cell lung cancer patients treatment.The growing evidences revealed that EGFR exon 19 and 21 mutation status in NSCLC patients was correlated with the outcome for EGFR-TKI treatment.In this study,two methods of Real-time PCR and DNA sequencing were compared to detected EGFR exon 19 and 21 mutations.Methods EGFR exon19 mutation del-E746-A750 and exon 21mutation L858R were detected by Real-time PCR and DNA sequencing in 103 NSCLC patients.Chi-square test was used to analyze the consistance.Results There was no significant difference between the two methods.However,Real-time PCR was more convenient and sensitive compared to DNA sequencing.Conclusion Real-time PCR was more suitable for clinical testing than DNA sequencing.