The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, Israel.
Viruses. 2020 Oct 15;12(10):1165. doi: 10.3390/v12101165.
Routine methods for virus detection in clinical specimens rely on a variety of sensitive methods, such as genetic, cell culture and immuno-based assays. It is imperative that the detection assays would be reliable, reproducible, sensitive and rapid. Isolation of viruses from clinical samples is crucial for deeper virus identification and analysis. Here we introduce a rapid cell-based assay for isolation and detection of viruses. As a proof of concept several model viruses including West Nile Virus (WNV), Modified Vaccinia Ankara (MVA) and Adenovirus were chosen. Suspended Vero cells were employed to capture the viruses following specific antibody labeling which enables their detection by flow cytometry and immuno-fluorescence microscopy assays. Using flow cytometry, a dose response analysis was performed in which 3.6e4 pfu/mL and 1e6 pfu/mL of MVA and WNV could be detected within two hours, respectively. When spiked to commercial pooled human serum, detection sensitivity was slightly reduced to 3e6 pfu/mL for WNV, but remained essentially the same for MVA. In conclusion, the study demonstrates a robust and rapid methodology for virus detection using flow cytometry and fluorescence microscopy. We propose that this proof of concept may prove useful in identifying future pathogens.
临床标本中病毒的常规检测方法依赖于多种敏感方法，如遗传、细胞培养和免疫测定。检测方法必须可靠、可重复、敏感和快速。从临床样本中分离病毒对于更深入的病毒鉴定和分析至关重要。在这里，我们介绍了一种用于病毒分离和检测的快速基于细胞的检测方法。作为概念验证，选择了几种模式病毒，包括西尼罗河病毒（WNV）、改良安卡拉痘苗病毒（MVA）和腺病毒。悬浮的 Vero 细胞被用于捕获经过特异性抗体标记的病毒，这使其能够通过流式细胞术和免疫荧光显微镜检测。通过流式细胞术进行剂量反应分析，可在两小时内分别检测到 3.6e4 pfu/mL 和 1e6 pfu/mL 的 MVA 和 WNV。当加入商业混合人血清中时，WNV 的检测灵敏度略有降低至 3e6 pfu/mL，但 MVA 的检测灵敏度基本保持不变。总之，该研究证明了使用流式细胞术和荧光显微镜进行病毒检测的强大而快速的方法。我们建议，这一概念验证可能有助于鉴定未来的病原体。
