Department of Chemical Engineering, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
J Virol Methods. 2012 Jun;182(1-2):27-36. doi: 10.1016/j.jviromet.2012.03.001. Epub 2012 Mar 8.
The increasing use of the baculovirus expression vector system (BEVS) has generated significant interest into techniques for quantifying baculovirus stocks. One method involves the use of quantitative real-time polymerase chain reaction (PCR). This study investigated simplifying baculovirus sample preparation for quantitative Real Time PCR to provide an alternative to current kit-based preparation methods. To achieve this goal, combinations of freeze/thaw cycles and Triton X-100 treatment were investigated. A treatment with only Triton X-100 was found to be sufficient to provide a simple, rapid and cheap alternative to kit-based preparation methods. This study also examined other factors such as primer choice to further examine the process of baculovirus quantitation by qPCR.
杆状病毒表达载体系统（BEVS）的使用越来越多，人们对定量杆状病毒的技术产生了浓厚的兴趣。一种方法涉及使用定量实时聚合酶链反应（PCR）。本研究旨在简化定量实时 PCR 用杆状病毒样本制备，以提供一种替代当前基于试剂盒的制备方法的方案。为了实现这一目标，研究了冻融循环和 Triton X-100 处理的组合。结果发现，仅用 Triton X-100 处理就足以提供一种简单、快速且廉价的替代试剂盒制备方法的方案。本研究还研究了其他因素，如引物选择，以进一步检查 qPCR 定量杆状病毒的过程。
