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

Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Food and Environmental Virology Springer Journals

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

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References (21)

Publisher
Springer Journals
Copyright
Copyright © 2011 by Springer Science + Business Media, LLC
Subject
Biomedicine; Chemistry/Food Science, general; Virology; Food Science
ISSN
1867-0334
eISSN
1867-0342
DOI
10.1007/s12560-011-9074-5
pmid
23412764
Publisher site
See Article on Publisher Site

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.

Journal

Food and Environmental VirologySpringer Journals

Published: Dec 17, 2011

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