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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 of 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 10~3 TO 10~9 virus particles per 200 muL of supernatant,and the results were closely correlated with titers detected from 50% tissue culture infectious doses (TCID_(50)) of baculovirus.This quantitative real-time PCR requires only 30 min to perform,and the entire titer determination can be accomplished within 1h 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：滴度测定是研究病毒的前提。但是，目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，简便的定量检测杆状病毒滴度的常规方法实时PCR。该方法基于扩增选定基因编码区中约150 bp的片段，发现PCR的定量范围为每200μL10〜3至10〜9个病毒颗粒上清液的滴度，结果与从杆状病毒的50％组织培养感染剂量（TCID_（50））检测到的滴度密切相关。此定量实时PCR只需30分钟即可完成，整个滴度测定可在不到30分钟的时间内完成1h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，可生成高精度数据，最重要的是非常快，因此肯定会将来可广泛用于杆状病毒的滴度测定。

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