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Rapid titer determination of baculovirus by quantitative real-time polymerase chain reaction

机译：定量实时聚合酶链反应快速滴度测定杆状病毒

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摘要

Titer determination is a prerequisite for the study of viruses. However, the current available methods are tedious and time-consuming. To improve the efficiency of titer determination, we have developed a rapid and simple method for the routine detection of baculovirus titers using a quantitative real-time PCR. This method is based on the amplification of approximately 150-bp fragments located in the coding regions of selected genes. The PCR was found to be quantitative in a range of 103 to 109 virus particles per 200 muL of supernatant, and the results were closely correlated with titers detected from 50% tissue culture infectious doses (TCID50) of baculovirus. This quantitative real-time PCR requires only 30 min to perform, and the entire titer determination can be accomplished within 1 h without the need for cell seeding or further virus dilution and infection. Because this technology is easy to operate, generates data with high precision, and most importantly is very quick, it will certainly be broadly applied for titer determination of baculoviruses in the future.
机译：确定滴度是研究病毒的前提。但是，当前可用的方法繁琐且耗时。为了提高滴度测定的效率，我们开发了一种快速简单的方法，用于使用定量实时PCR常规检测杆状病毒滴度。该方法基于位于所选基因编码区的约150 bp片段的扩增。发现该PCR在每200μL上清液103至109个病毒颗粒的范围内是定量的，并且该结果与从杆状病毒的50％组织培养物感染剂量（TCID50）检测到的效价密切相关。这种实时定量PCR只需30分钟即可完成，整个滴定度测定可在1小时内完成，而无需细胞接种或进一步的病毒稀释和感染。由于该技术易于操作，生成高精度数据，并且最重要的是非常快，因此将来肯定会广泛应用于杆状病毒的滴度测定。

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