Development of High-Content Imaging Assays for Lethal Viral Pathogens
Development of High-Content Imaging Assays for Lethal Viral Pathogens
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DOI：
10.1177/1087057110374357
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发表时间：
2010-08-01
影响因子：
--
通讯作者：
Bavari, Sina
中科院分区：
文献类型：
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作者：
Panchal, Rekha G.;Kota, Krishna P.;Bavari, Sina
埃博拉病毒（EBOV）和马尔堡病毒（MARV）等丝状病毒是单链负感RNA病毒，可引起死亡率高的急性出血热。目前，还没有获得许可的疫苗或治疗方法来对抗人类丝状病毒感染。开发高通量/高含量的初级筛选分析，然后使用低通量的传统斑块或实时PCR分析进行验证，将极大地有助于发现新的抗病毒治疗方法。具体来说，高含量成像技术越来越多地应用于药物的初级筛选。在这项研究中，作者描述了在优化基于图像采集和分析高致病性丝状病毒埃博拉和马尔堡的生物测定时遇到的挑战。在开发这些检测方法的过程中，评估了许多生物学和成像相关的变量，如镀膜密度、感染的多重性、每孔扫描的场数、荧光强度和分析的细胞数量。此外，作者证明了与单细胞数据统计分析相关的好处，以解释病毒抗原染色模式的亚细胞定位和全细胞整合强度的异质性。总之，它们表明基于图像的方法是鉴定高致病性病毒抗病毒化合物的有力筛选工具。（Journal of biomolmolecular Screening 2010:755-765）
Filoviruses such as Ebola (EBOV) and Marburg (MARV) are single-stranded negative sense RNA viruses that cause acute hemorrhagic fever with high mortality rates. Currently, there are no licensed vaccines or therapeutics to counter filovirus infections in humans. The development of higher throughput/high-content primary screening assays followed by validation using the low-throughput traditional plaque or real-time PCR assays will greatly aid efforts toward the discovery of novel antiviral therapeutics. Specifically, high-content imaging technology is increasingly being applied for primary drug screening. In this study, the authors describe the challenges encountered when optimizing bioassays based on image acquisition and analyses for the highly pathogenic filoviruses Ebola and Marburg. A number of biological and imaging-related variables such as plating density, multiplicity of infection, the number of fields scanned per well, fluorescence intensity, and the cell number analyzed were evaluated during the development of these assays. Furthermore, the authors demonstrate the benefits related to the statistical analyses of single-cell data to account for heterogeneity in the subcellular localization and whole-cell integrated intensity of the viral antigen staining pattern. In conclusion, they show that image-based methods represent powerful screening tools for identifying antiviral compounds for highly pathogenic viruses. (Journal of Biomolecular Screening 2010:755-765)