中文摘要：

目的通过对合并胸腔积液的非小细胞肺癌（NSCLC）患者胸腔积液KRAS基因的检测,建立非组织来源KRAS检测的替代方法,探讨KRAS突变情况及主要临床病理特征。方法收集30例NSCLC胸腔积液标本,用焦磷酸测序法和锁核酸探针-桑格测序法（LNA-PCR sanger）检测各标本中KRAS突变情况,统计分析KRAS突变与临床病理特征之间的关系。结果焦磷酸测序法检出3例突变,而LNA-PCR sanger测序法则检出6例突变。6例突变中有4例位于第2外显子第12号密码子,2例位于第2外显子第14号密码子。统计学分析KRAS突变与性别、吸烟史、年龄、病理类型及体力状况（PS）评分无关。结论 LNA-PCR sanger测序和焦磷酸测序法均可用于胸腔积液标本KRAS突变检测,可为NSCLC基因检测提供良好的非组织标本替代方法。

英文摘要：

Objective KRAS mutations are associated with the epidermal growth factor receptor tyrosine kinase inhibitor（EGFR-TKI）-resistance,which has become the bottle-neck problem in molecular targeted therapy of non-small cell lung cancer（NSCLC）.In this study,we detected KRAS mutations in pleural effusion of NSCLC patients,in order to investigate its mutant status and the major clinical pathological characteristics,and establish vicarious method for KRAS-mutation detection to non-histological sample.Methods The pleural effusions of 30 patients with NSCLC were collected.The mutant status of KRAS were analyzed by the pyrosequencing and the locked nucleic acids probes-PCR（LNA-PCR） sanger sequence.The relationships of mutant status and the major clinical pathological characteristics were analyzed.Results The KRAS mutation rate in pleural effusion of 30 cases was 10%（3/30） by pyrosequencing and 20%（6/30）by LNA-PCR sanger sequencing,respectively.Among 6 cases of KRAS mutation,4 cases were located in code 12,and 2 cases were located in code 14.KRAS mutation did not relate with gender,smoking,histological classification and PS score.Conclusion KRAS mutations in pleural effusion could be detected by LNA-PCR sanger sequencing and pyrosequencing,which might become favourable methods for NSCLC gene detection.