HBV-DNA定量标准品的构建

Construction of plasmid standards for quantitation of HBV-DNA

目的建立一种简便快速的DNA定量标准品的制备方法。方法PCR扩增目的片段,与T载体连接,转化感受态大肠杆菌DH5α,阳性克隆进行测序证实。结果标准品建立的标准曲线有较大的线性范围,批内和批间重复性也较好,且可长期稳定保存。结论使用质粒标准品对样本中病毒载量进行测定是一种可行的办法。

Objective To establish a simple and quick method for preparation of DNA quantitative standard substance. Methods Target genes amplified with T vector were transformed into competent E.Coli. DH5 α, and identified by real-time fluorescence quantitative PCR assay. Results Standard curve created by DNA plasmid standards had wide detection range, and the intraexperimental and interexperimental variability was also fine. Conclusion It is viable that the plasmid standards can be used for virus loads detection.