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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。
Objective: To establish a fluorescent quantitative PCR (FQ-PCR) method for c-erbB-2 gene in breast cancer. METHODS: The c-erbB-2 gene in breast cancer cells was recombined with the plasmid PGEM-Teasy vector and transformed into E. coli DH5α to obtain the cloned standard template of c-erbB-2 gene. The ABI PRISM7700PCR instrument was used to detect the FQ-PCR amplification product to make a standard curve to detect the content of c-erbB-2 in the unknown sample. RESULTS: The FQ-PCR products showed a “S” kinetic curve; there was a strict linear relationship between the ct (cycle threshold) and the logarithm of the initial template copy number in the PCR system, indicating that the quantitative FQ-PCR was accurate Sex. Conclusion: FQ-PCR is a rapid, simple, sensitive and accurate method for quantifying c-erbB-2 gene.