【摘要】 目的:建立乳腺癌中c-erbB-2基因的荧光定量PCR(FQ-PCR)方法。方法:乳腺癌细胞中的c-erbB-2基因与质粒PGEM-Teasy vector重组,转化大肠杆菌E.coliDH5α,获得克隆的c-erbB-2基因标准模板。用ABI PRISM7700PCR仪检测FQ-PCR扩增产物制成标准曲线来检测未知标本中c-erbB-2的含量。结果:FQ-PCR扩增产物呈“S”形动力学曲线;ct(循环阈值)与PCR体系中起始模板拷贝数的对数值之间存在严格的线性关系,显示了FQ-PCR定量的准确性。结论:FQ-PCR是一种快速、简便、灵敏、准确的定量c-erbB-2基因的方法。更多还原

【Abstract】 AIM: To establish a fluorogenic quantitative polymerase chain reaction (FQ-PCR) method for the routine examination of c-erbB-2 gene expression in breast cancer. METHODS: The c-erbB-2 standard gene was obtained by in vitro amplification of cloned c-erbB-2 fragment in plasmid PGEM-T easy vector. FQ-PCR product was detected by using a 7700 ABI PRISM sequence detector system and c-erbB-2 standard curve was obtained to quantity c-erbB-2 in unknown samples. RESULTS: “S” kinetics curve of FQ-PCR amplification was generated by relating the fluorescence signal intensity (△Rn) to the cycle number. The standard curve of c-erbB-2 was constructed by the linear relationship between the cycle threshold (ct) and the log of starting copy number. The high correlation (0.999) revealed the reliability of FQ-PCR. CONCLUSION: The FQ-PCR is a rapid, sensitive, reliable method for quantity of c-erbB-2 gene expression.更多还原