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International Conference on SARS - one year after the (first) outbreak

08. - 11.05.2004, Lübeck

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A tetrazolium-based colorimetric cell culture assay for the identification of SARS coronavirus inhibitors

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  • corresponding author presenting/speaker Els Keyaerts - Rega Institute for Medical Research, KULeuven, Leuven, Belgium
  • Leen Vijgen - Rega Institute for Medical Research, KULeuven, Leuven, Belgium
  • Johan Neyts - Rega Institute for Medical Research, KULeuven, Leuven, Belgium
  • Erik de Clercq - Rega Institute for Medical Research, KULeuven, Leuven, Belgium
  • Jan Balzarini - Rega Institute for Medical Research, KULeuven, Leuven, Belgium
  • Marc van Ranst - Rega Institute for Medical Research, KULeuven, Leuven, Belgium; U.Z. Leuven, Leuven, Belgium

International Conference on SARS - one year after the (first) outbreak. Lübeck, 08.-11.05.2004. Düsseldorf, Köln: German Medical Science; 2004. Doc04sarsP9.05

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/sars2004/04sars127.shtml

Published: May 26, 2004

© 2004 Keyaerts et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

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Severe acute respiratory syndrome (SARS) has recently emerged as a new severe human disease, resulting globally in 774 deaths from 8098 reported probable cases. A novel member of the Coronaviridae family has been identified as the causative agent of this pulmonary disease. Although the initial global outbreak of SARS appears to have been successfully contained, SARS will remain a serious concern while there continues to be no suitable vaccine or effective drug treatment. A colorimetric assay based on the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) into a water soluble formazan product which can be directly quantified using a microtiter ELISA reader, has been developed for SARS coronavirus strain Frankfurt 1 drugsusceptibility testing. Optimal conditions were determined and the standard routine assay was calibrated with a viral input of 100 CCID50 and a density of 10.000 cells per well in a 96-well microtiter plate for an incubation period of 3 days. Interferon β was used as a positive control to validate the assay. The effective IC50 concentration value obtained with interferon β in the present assay was in agreement with interferon β activity results published by others. This method presents the advantage of being rapid, reliable, reproducible, and convenient for high throughput screening capacity in a stringent P3 biosafety environment.