中文摘要：

目的 比较临床常用的两种方法检测晚期非小细胞肺癌（NSCLC）表皮生长因子受体（EGFR）基因突变状态结果的差异,探讨福尔马林固定石蜡包埋（FFPE）组织切片标本的不同保存方式对DNA数量、质量及对检测结果的影响.方法 收集中国医学科学院肿瘤医院2010年11月至2011年8月晚期NSCLC FFPE组织切片标本150份,分别采用楔形探针扩增阻滞突变系统（ScorpionsARMS）和直接测序法检测EGFR第18、19、20、21号外显子突变状态,分析两种方法检测EGFR基因突变结果的差异,对结果不一致病例收集临床治疗情况及生存数据并进行分析.对其中30份FFPE标本提取DNA并保存于-20℃3年,同样的30份FFPE标本按常规方法保存3年后再提取DNA,比较两种保存方式对DNA的数量、质量及对检测结果的影响.结果 Scorpions ARMS法检测EGFR基因突变的成功率（100％）高于直接测序法（87.3％）,差异有统计学意义（χ^2=20.28,P＜0.05）.两种方法检测EGFR基因突变的一致率为84.7％,一致性较好（Kappa=0.738,P＜0.001）.分析结果不一致病例中5例接受靶向药物治疗后患者的生存情况,与ARMS检测结果一致性相对较好.新鲜FFPE标本提取并贮存于-20℃、3年的DNA与FFPE标本按常规方法保存3年后提取的DNA相比,后者质量相对较差,片段化程度更严重,但两种保存方法获取的DNA的EGFR基因突变检测结果均一致.结论 Scorpions ARMS与直接测序法联合检测结果更为全面、可靠.检测结果不确定的患者应以临床治疗及生存资料支持诊断.FFPE标本提取DNA后低温贮存,更有利于DNA完整性的保存.

英文摘要：

Objective To compare two different methods in detecting epidermal growth factor receptor（EGFR） mutation status of non-small cell lung cancer （NSCLC）,and investing the influences of different conserving methods in DNA quantity,quality and detecting results of formalin-fixed paraffin embedded （FFPE） tissue samples.Methods One hundred and fifty FFPE samples of advanced NSCLC were collected in Cancer Institute/Hospital,Chinese Academy of Medical Sciences from November 2010 to August 2011.Both scorpions amplification refractory mutation system（scorpions ARMS）and direct sequencing were used to detect EGFR mutation in exon 18,19,20 and 21,together analyzed the differences between these two methods.Samples with inconsistent results were collected clinical treatment and survival information for further analysis.Extracted DNA from 30 FFPE samples and conserved at-20 ℃ for 3 years,meanwhile the same 30 FFPE samples DNA were extracted after conserving for 3 years.The DNA quantity、quality and testing results of the two conserving methods were compared.Results The detection success rate of Scorpions ARMS （100%） was higher than direct sequencing （87.3%）,with statistical significance （χ^2 =20.28,P 〈 0.05）.The consistent rate of the two methods was 84.7%,and there were no significant differences between these two methods,with a high consistence （Kappa =0.738,P 〈 0.001）.The clinical treatment and survival status of 5 patients with different testing results were analysed,which were consistent with ARMS testing results.The quality of DNA extracted from FFPE tissues which were conserved for 3 years according to the conventional way was worse and with more fragments compared with DNA conserved at -20 ℃ for 3 years,but the EGFR mutation detecting results of the DNA acquired from the two conserved ways were consistent.Conclusions Combination of Scorpions ARMS and direct sequencing could make the detecting result more comprehensive and reliable.Clinical treatment and survival data may provide di