Development of high-content imaging assays for lethal viral pathogens

Journal of Biomolecular Screening
Rekha G PanchalSina Bavari

Abstract

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 inten...Continue Reading

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Citations

Nov 8, 2011·Biosecurity and Bioterrorism : Biodefense Strategy, Practice, and Science·Aruna SampathJoseph C Larsen
Oct 19, 2011·The Journal of Infectious Diseases·Kristina Maria SchmidtElke Mühlberger
Jan 29, 2011·Journal of Virology·Arnab BasuTerry L Bowlin
Sep 27, 2012·Viruses·Gianluca PegoraroRekha G Panchal
Nov 22, 2012·Viruses·Brian M FriedrichGene G Olinger
Aug 10, 2012·Expert Opinion on Drug Discovery·Amy C ShurtleffSina Bavari
Oct 27, 2011·Antiviral Research·Rekha G PanchalSina Bavari
Jul 10, 2015·ACS Infectious Diseases·Peter B MadridMary J Tanga
Jun 12, 2015·ACS Infectious Diseases·Života SelakovićBogdan A Šolaja
Jun 21, 2011·Current Opinion in Chemical Biology·Priscille Brodin, Thierry Christophe
Aug 30, 2014·Expert Review of Anti-infective Therapy·Thomas Hoenen, Heinz Feldmann
Nov 9, 2018·European Journal of Medicinal Chemistry·Života SelakovićBogdan A Šolaja

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Methods Mentioned

BETA
enzyme-linked immunosorbent assay
optical microscopy

Software Mentioned

Acapella
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Excel
Opera
SAS
Opera Evotec

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