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Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR

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Abstract

The discrimination of infectious and inactivated viruses remains a key obstacle when using quantitative RT-PCR (RT-qPCR) to quantify enteric viruses. In this study, propidium monoazide (PMA) and RNase pretreatments were evaluated for the detection and quantification of infectious hepatitis A virus (HAV). For thermally inactivated HAV, PMA treatment was more effective than RNase treatment for differentiating infectious and inactivated viruses, with HAV titers reduced by more than 2.4 log10 units. Results showed that combining 50 μM of PMA and RT-qPCR selectively quantify infectious HAV in media suspensions. Therefore, PMA treatment previous to RT-qPCR detection is a promising alternative to assess HAV infectivity.

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Acknowledgements

This study was partially supported by grant AGL2009-08603 from the Spanish Ministry of Science and Innovation and grant ACOMP/2010/279 from the Generalitat Valenciana. G. Sánchez is the recipient of a JAE post-doc grant from the “Consejo Superior de Investigaciones Científicas” (CSIC). We thank Loles Rodrigo for her scientific and technical support on HPP and Sonia Gonzalez for technical support.

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Authors and Affiliations

  1. Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos, Avda. Agustín Escardino, 7. Paterna, Valencia, Spain

    Gloria Sánchez

  2. Department of Microbiology and Ecology, University of Valencia, Valencia, Spain

    Patricia Elizaquível & Rosa Aznar

  3. Institute of Agrochemistry and Food Technology (IATA), Spanish Council for Scientific Research (CSIC), Valencia, Spain

    Rosa Aznar

Authors
  1. Gloria Sánchez
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  2. Patricia Elizaquível
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  3. Rosa Aznar
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Correspondence to Gloria Sánchez.

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Sánchez, G., Elizaquível, P. & Aznar, R. Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR. Food Environ Virol 4, 21–25 (2012). https://doi.org/10.1007/s12560-011-9074-5

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  • DOI: https://doi.org/10.1007/s12560-011-9074-5

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