Real-time monitoring of flavivirus induced cytopathogenesis using cell electric impedance technology

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• 作者：Ying Fang^a ; Peifang Ye^b ; Xiaobo Wang^b ; Xiao Xu^b ; William Reisen^a ; ^{wkreisen@ucdavis.edu}
• 关键词：Real-time cell analysis system ; Viral cytopathogenesis ; West Nile virus ; St. Louis encephalitis virus ; Impedance ; Neutralization assay
• 刊名：Journal of Virological Methods
• 出版年：2011
• 出版时间：May 2011
• 年：2011
• 卷：173
• 期：2
• 页码：251-258
• 全文大小：698 K

文摘

A real-time cell analysis (RTCA) system based on cell-substrate electric impedance technology was used to monitor cytopathic effects (CPE) in Vero cell cultures infected with West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) at infectious doses ranging from 10¹ to 10⁶ plaque forming units (PFU) of virus. A kinetic parameter characterizing virus-induced CPE, CIT₅₀ or the time to 50 % decrease in cell impedance, was inversely proportional to virus infectious dose. In WNV-infected cells, the onset and rate of CPE was earlier and faster than in SLEV-infected cells, which was consistent with viral cytolytic activity. A mathematical model simulating impedance-based CPE kinetic curves indicated that the replication rate of WNV was about 3 times faster than SLEV. The RTCA system also was used for quantifying the level of cell protection by specific neutralizing antibodies against WNV and SLEV. The onset of WNV or SLEV-induced CPE was delayed in the presence of specific anti-sera, and this delay in the CIT₅₀ was well correlated with the titer of the neutralizing antibody as measured independently by plaque reduction neutralization tests (PRNT). The RTCA system provided a high throughput and quantitative method for real-time monitoring viral growth in cell culture and its inhibition by neutralizing antibodies.