【摘要】 目的建立一种荧光定量逆转录聚合酶链反应(FQ RT-PCR)检测肺癌患者外周血hnRNP B1mRNA表达的相对定量方法。方法以hnRNP B1为待测基因,以β-actin为参照进行检测,建立标准曲线,确定实时RT-PCR的扩增效率;优化反应条件,使扩增效率接近100%,检测待检标本hnRNP B1和β-actin的循环阈值(Ct)。结果该法检测的最低拷贝数为103,线性范围为103～107拷贝,批内和批间变异系数(CV)分别为3.4%和11.3%。结论该方法具有灵敏、特异、数据处理简便可靠等特点,为其他肿瘤患者外周血中肿瘤细胞的检测提供了方法学启示。更多还原

【Abstract】 Objective To develop a fluorescence quantitative reverse transcriptase polymerase chain reaction(FQ RT-PCR)method for detecting hnRNP B1 mRNA in peripheral blood of lung cacinoma patients.Methods hnRNP B1 was used as target gene,and β-actin as reference.Stand curves were established to measure the amplification efficiency of hnRNP B1 and β-actin.By changing reaction conditions,the amplification efficiency of two gene was nearly 100%.Ct values of hnRNP B1 and β-actin were measured in samples stimultaneously.Results The method could detect as low as 103 copies and the linear range was 103～107 copies,the intra-assay variation was 3.4%,the inter-assay variation was 11.8%.Conclusions The method is a sensitive,specific procedure with simple and reliable data analysis,which providing a promising method for the detection of follow-up patients of different kinds of cacinoma.更多还原