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使用电细胞基质阻抗传感作为实时定量甲型流感病毒感染的MDCK细胞中细胞病变效应的工具。

Use of electric cell-substrate impedance sensing as a tool for quantifying cytopathic effect in influenza A virus infected MDCK cells in real-time.

作者信息

McCoy Morgan H, Wang Eugenia

机构信息

University of Louisville, Department of Biochemistry and Molecular Biology, Gheens Center on Aging, Delia Baxter Building II, rm 102, 580 South Preston Street, Louisville, KY 40202, USA.

出版信息

J Virol Methods. 2005 Dec;130(1-2):157-61. doi: 10.1016/j.jviromet.2005.06.023. Epub 2005 Aug 10.

Abstract

Resulting from its subjective nature, cytopathic effect (CPE) due to virus infection in cell culture has long been difficult to quantify. This report illustrates the use of electric cell-substrate impedance sensing (ECIS) for monitoring the progression of CPE due to influenza A virus infection. ECIS monitors the impedance of a non-invasive ac current flowing through cell culture medium by gold film electrodes placed on the surface of the culture dish. As cultured cells attach and spread onto the electrodes, the current is impeded proportional to the number of attached cells, the number of tight junctions between cells and the shortness in distance between the cells and the substratum. In the case at hand, a healthy monolayer of cells was insulted with influenza A virus infection and exhibited a characteristic rounded cell morphology and cell detachment. These effects resulted in reduced impedance, which was monitored with ECIS. Since data obtained through ECIS are both quantitative and in real-time, it was possible to monitor continuously cell behavior during infection. This, in turn, allowed for a more detailed and comprehensive data set to analyze. More importantly, through ammonium chloride treatment of cells, it was also shown that ECIS may be exploited to examine a treatment's effect on the reduction of resistance because of its antiviral activity. Thus, ECIS may be a powerful approach for screening antiviral compounds quantitatively in a real-time fashion.

摘要

由于其主观性,细胞培养中病毒感染引起的细胞病变效应(CPE)长期以来一直难以量化。本报告阐述了利用电场细胞-基质阻抗传感(ECIS)监测甲型流感病毒感染导致的CPE进展情况。ECIS通过置于培养皿表面的金膜电极监测流过细胞培养基的无创交流电流的阻抗。随着培养细胞附着并铺展在电极上,电流受到的阻碍与附着细胞的数量、细胞间紧密连接的数量以及细胞与基质之间距离的缩短成正比。在当前案例中,健康的单层细胞受到甲型流感病毒感染,呈现出特征性的细胞变圆形态和细胞脱离。这些效应导致阻抗降低,通过ECIS进行监测。由于通过ECIS获得的数据兼具定量性和实时性,因此有可能在感染期间持续监测细胞行为。这反过来又能得到更详细、更全面的数据集以供分析。更重要的是,通过对细胞进行氯化铵处理,还表明由于其抗病毒活性,ECIS可用于检测一种治疗方法对降低耐药性的效果。因此,ECIS可能是一种以实时方式定量筛选抗病毒化合物的有力方法。

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