Original Paper

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 SánchezAffiliated withDepartamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos Email author 
  • , Patricia ElizaquívelAffiliated withDepartment of Microbiology and Ecology, University of Valencia
  • , Rosa AznarAffiliated withDepartment of Microbiology and Ecology, University of ValenciaInstitute of Agrochemistry and Food Technology (IATA), Spanish Council for Scientific Research (CSIC)

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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.

Keywords

Hepatitis A virus Quantitative PCR Infectivity Propidium monoazide