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

被引:0
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作者
Gloria Sánchez
Patricia Elizaquível
Rosa Aznar
机构
[1] Instituto de Agroquímica y Tecnología de Alimentos,Departamento de Biotecnología
[2] University of Valencia,Department of Microbiology and Ecology
[3] Institute of Agrochemistry and Food Technology (IATA),undefined
[4] Spanish Council for Scientific Research (CSIC),undefined
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Hepatitis A virus; Quantitative PCR; Infectivity; Propidium monoazide;
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摘要
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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页码:21 / 25
页数:4
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