【摘要】 研究应用ARMS-PCR技术分别为表皮生长因子受体(epidermal growth factor receptor,EGFR)基因19外显子缺失和L858R点突变建立3种不同长度扩增子定量检测方法。实验采用不同长度扩增子检测95例非小细胞肺癌(non-small cell lung cancer,NSCLC)患者新鲜冰冻肿瘤组织样本和108例患者石蜡切片样本(formalin-fixation and paraffin-embedding,FFPE)19外显子缺失和L858R突变。结果显示携带突变的FFPE样本中,长度最短扩增子(RDEL-1和RL858R-1)检测样本突变结果显著优于其他2种扩增子(P<0.001;P<0.001);新鲜冰冻肿瘤组织样本不同扩增子检测结果无显著性差异(19外显子缺失:P=0.270;L858R:P=0.249);不同扩增子对野生型(wild type,WT)样本具有很高的检测特异性。研究结果表明试验中采用的几种不同长度扩增子ARMS-PCR方法可适用于新鲜冰冻肿瘤组织EGFR突变检测,但FFPE样本更优选最短的扩增子(RDEL-1和RL858R-1)检测方法。更多还原

【Abstract】 In this study,mutations of epidermal growth factor receptor(EGFR),including exon 19 deletion and point mutation L858 Rwere quantitatively detected with AMRS-PCR by using three amplicons with different lengths.The tumor tissue samples came from patients with non-small cell lung cancer(NSCLC)consisting of 95 fresh frozen samples and 108formalin-fixation and paraffin-embedding(FFPE)samples.The results showed that for the FFPE samples,the shortest amplicons RDEL-1and RL858 Rwere showed significantly better performance than the other amplicons on the detection of mutations in FFPE samples(P < 0.001,P< 0.001),whereas for the fresh frozen tissue,amplicons with different lengths did not show significant differences(exon 19 deletion,P=0.270;L858R,P=0.249).All amplicons had a high specificity for wild type specimens.Altogether,these data demonstrate that using ARMS-PCR method,different lengths of amplicons are applicable for determining EGFR mutant type in fresh frozen tissue;however,the amplicons with shorter lengths,RDEL-1and RL858R-1,seem to be a better choice for FFPE samples.更多还原