Department of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea.
Department of Mechanical Engineering, Yonsei University, Seoul 03722, Republic of Korea; Institute of Engineering Research, Yonsei University, Seoul 03722, Republic of Korea.
J Hazard Mater. 2024 Nov 5;479:135544. doi: 10.1016/j.jhazmat.2024.135544. Epub 2024 Aug 22.
Accurate airborne virus monitoring is important for preventing the spread of infectious diseases. Although standard reverse transcription-quantitative polymerase chain reaction (RT-qPCR) can efficiently detect viral ribonucleic acid (RNA), it cannot determine whether the RNA is associated with active (infectious) or inactive (non-infectious) viruses. Plaque assay is the gold standard for determining viral infectivity but is laborious and time-consuming. This study explored the viral infectivity of H1N1 influenza virus and human coronavirus (HCoV-229E) using capsid integrity RT-qPCR, where virus samples were pretreated with reagents penetrating viruses with damaged capsids, impeding amplification by binding to their RNA. Therefore, the amplified signals corresponded solely to active viruses with undamaged capsids. Propidium monoazide (PMA) and platinum (IV) chloride (PtCl) were used to investigate the effects of reagent concentration. Feasibility tests revealed that PtCl was more efficient than PMA, with optimal concentrations of 125-250 μM and 250-500 μM for H1N1 influenza virus and HCoV-229E, respectively. The results of percentage of active virus showed that capsid integrity RT-qPCR provided a trend similar to that of plaque assay, indicating an accurate measure of viral infectivity. Virus sampling in the laboratory and field highlighted the precision of this methodology for determining viral infectivity. Therefore, this methodology enables rapid and accurate detection of infectious airborne H1N1 influenza virus and HCoV-229E, allowing swift response to outbreaks.
准确的空气传播病毒监测对于预防传染病的传播非常重要。虽然标准的逆转录-定量聚合酶链反应（RT-qPCR）可以有效地检测病毒的核糖核酸（RNA），但它无法确定 RNA 是否与活性（感染性）或非活性（非感染性）病毒有关。噬斑测定法是确定病毒感染力的金标准，但繁琐且耗时。本研究使用衣壳完整性 RT-qPCR 探索了 H1N1 流感病毒和人类冠状病毒（HCoV-229E）的病毒感染力，其中病毒样本用穿透有损伤衣壳的病毒的试剂预处理，通过与它们的 RNA 结合来阻碍扩增。因此，扩增信号仅对应于未受损衣壳的活性病毒。吖啶橙（PMA）和四价铂（IV）氯化物（PtCl）用于研究试剂浓度的影响。可行性测试表明，PtCl 比 PMA 更有效，H1N1 流感病毒和 HCoV-229E 的最佳浓度分别为 125-250 μM 和 250-500 μM。活性病毒百分比的结果表明，衣壳完整性 RT-qPCR 提供了与噬斑测定法相似的趋势，表明这是一种准确测量病毒感染力的方法。实验室和现场的病毒采样突出了这种方法确定病毒感染力的精确性。因此，这种方法能够快速准确地检测空气中具有传染性的 H1N1 流感病毒和 HCoV-229E，从而迅速应对疫情爆发。
