细胞培养作为定量检测活病毒的金标准,耗时长,通常需要数天到数周,光敏染料叠氮溴化丙锭联合qPCR(PMA-qPCR)作为一种新型定量检测病毒感染性的分析工具,具有快速便捷的优点PMA预处理待测样品,选择性渗入灭活病毒衣壳,结合核酸后抑制其扩增,使得仅活病毒核酸可以进行qPCR本研究就PMA-qPCR检测活病毒的机理、灭活病毒的方法进行分析讨论,并比较不同方案检测病毒感染性的差异性。
Plaque experiment and median tissue culture infective dose ( TCID50 ), both based on cell culture, are wildly used as the gold standard in discrimination of infectious viruses, but they are time consuming. Propidium monoaz- ide (PMA) combined with quantitative PCR (qPCR) is a new method of discrimination between infectious virus and noninfectious viruses. Samples are given PMA pretreatment before qPCR. PMA infiltrates damaged capsid selectively, conbines the nucleic acid and inhibits its transcription and amplification. This review elaborates the mechanism of PMA- qPCR in detecting infectious viruses, summarizes the experiment condition of PMA pretreatment, and discusses the effi- ciency of different approaches in discrimination of infectious virus.