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A Colorimetric-Based Accurate Method for the Determination of Enterovirus 71 Titer

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Abstract

The 50 % tissue culture infectious dose (TCID50) is still one of the most commonly used techniques for estimating virus titers. However, the traditional TCID50 assay is time consuming, susceptible to subjective errors and generates only quantal data. Here, we describe a colorimetric-based approach for the titration of Enterovirus 71 (EV71) using a modified method for making virus dilutions. In summary, the titration of EV71 using MTT or MTS staining with a modified virus dilution method decreased the time of the assay and eliminated the subjectivity of observational results, improving accuracy, reproducibility and reliability of virus titration, in comparison with the conventional TCID50 approach (p < 0.01). In addition, the results provided evidence that there was better correlation between a plaquing assay and our approach when compared to the traditional TCID50 approach. This increased accuracy also improved the ability to predict the number of virus plaque forming units present in a solution. These improvements could be of use for any virological experimentation, where a quick accurate titration of a virus capable of causing cell destruction is required or a sensible estimation of the number of viral plaques based on TCID50 of a virus is desired.

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Acknowledgments

We wish to thank Professor Peter C. McMinn (Central Clinical School, University of Sydney, Australia) for providing the cloned virus and the cells and Dr Julian Druce (Polio reference laboratory, Victorian Infectious Disease Reference Laboratory, Parkville, VIC, Australia) for providing us with the clinical isolate used. We also thank Dr. Jillian Shaw for reading the manuscript.

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  1. Hamid Reza Pourianfar

    Present address: Iranian Academic Centre for Education, Culture and Research (ACECR)-Mashhad Branch, 91775-1376, Mashhad, Iran

Authors and Affiliations

  1. Environment and Biotechnology Centre, Swinburne University of Technology, Hawthorn, VIC, 3122, Australia

    Hamid Reza Pourianfar, Arman Javadi & Lara Grollo

Authors
  1. Hamid Reza Pourianfar
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  2. Arman Javadi
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  3. Lara Grollo
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Correspondence to Lara Grollo.

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Pourianfar, H.R., Javadi, A. & Grollo, L. A Colorimetric-Based Accurate Method for the Determination of Enterovirus 71 Titer. Indian J. Virol. 23, 303–310 (2012). https://doi.org/10.1007/s13337-012-0105-0

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  • DOI: https://doi.org/10.1007/s13337-012-0105-0

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