Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR |
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| Authors: | Gloria Sánchez Patricia Elizaquível Rosa Aznar |
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| Institution: | 1.Departamento de Biotecnología,Instituto de Agroquímica y Tecnología de Alimentos,Valencia,Spain;2.Department of Microbiology and Ecology,University of Valencia,Valencia,Spain;3.Institute of Agrochemistry and Food Technology (IATA), Spanish Council for Scientific Research (CSIC),Valencia,Spain |
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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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