Food and Environmental Virology

, Volume 4, Issue 1, pp 21–25

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

  • Gloria Sánchez
  • Patricia Elizaquível
  • Rosa Aznar
Original Paper

DOI: 10.1007/s12560-011-9074-5

Cite this article as:
Sánchez, G., Elizaquível, P. & Aznar, R. Food Environ Virol (2012) 4: 21. doi:10.1007/s12560-011-9074-5

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.

Keywords

Hepatitis A virus Quantitative PCR Infectivity Propidium monoazide 

Copyright information

© Springer Science + Business Media, LLC 2011

Authors and Affiliations

  • Gloria Sánchez
    • 1
  • Patricia Elizaquível
    • 2
  • Rosa Aznar
    • 2
    • 3
  1. 1.Departamento de BiotecnologíaInstituto de Agroquímica y Tecnología de AlimentosValenciaSpain
  2. 2.Department of Microbiology and EcologyUniversity of ValenciaValenciaSpain
  3. 3.Institute of Agrochemistry and Food Technology (IATA), Spanish Council for Scientific Research (CSIC)ValenciaSpain

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