Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles
Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles
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DOI：
10.1002/bit.21177
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发表时间：
2007-03-01
影响因子：
3.8
通讯作者：
King, Linda A.
中科院分区：
文献类型：
--
作者：
Hitchman, Richard B.;Siaterli, Evangelia A.;King, Linda A.
我们描述了使用定量聚合酶链式反应(QPCR)来滴定重组杆状病毒。根据gp64设计了定制的引物和探针，并用于根据先前滴定的杆状病毒种群的稀释度计算QPCR产生的滴度的标准曲线。每种稀释度都通过斑块分析和定量聚合酶链式反应进行滴定；在C-T和每毫升斑块形成单位之间产生一致且可重复的反关系。12株重组病毒的QPCR滴度与空斑滴度无显著差异，证明该方法是一种快速、准确的杆状病毒滴度检测方法。(C)2006年威利期刊公司。
We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay and QPCR; producing a consistent and reproducible inverse relationship between C-T and plaque forming units per milliliter. No significant difference was observed between titers produced by QPCR and plaque assay for 12 recombinant viruses, confirming the validity of this technique as a rapid and accurate method of baculovirus titration. (c) 2006 Wiley Periodicals, Inc.