Propidium monoazide reverse transcriptase PCR and RT-qPCR for detecting infectious enterovirus and norovirus

Journal of Virological Methods
Mohammad R KarimSandhya Parshionikar

Abstract

Presently there is no established cell line or small animal model that allows for the detection of infectious human norovirus. Current methods based on RT-PCR and RT-qPCR detect both infectious and non-infectious virus and thus the conclusions that may be drawn regarding the public health significance of positive findings are limited. In this study, PMA RT-PCR and RT-qPCR assays were evaluated for selective detection of infectious poliovirus, murine norovirus (MNV-1), and Norwalk virus. Viruses were inactivated using heat, chlorine, and ultraviolet light (UV). Infectious and non-infectious viruses were treated with PMA before RT-PCR and RT-qPCR. PMA RT-PCR was able to differentiate selectively between infectious and heat and chlorine inactivated poliovirus. PMA RT-PCR was able to differentiate selectively between infectious and noninfectious murine norovirus only when inactivated by chlorine. However, PMA RT-PCR could not differentiate infectious Norwalk virus from virus particles rendered non-infectious by any treatment. PMA RT-PCR assay was not able to differentiate between infectious and UV inactivated viruses suggesting that viral capsid damage may be necessary for PMA to enter and bind to the viral genome. PMA RT-PCR on na...Continue Reading

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Citations

Sep 10, 2017·Applied and Environmental Microbiology·Nathan DunkinKellogg J Schwab
Jun 25, 2019·Transboundary and Emerging Diseases·Ravo M RazafimahefaEtienne Thiry
Mar 7, 2017·Shokuhin eiseigaku zasshi. Journal of the Food Hygienic Society of Japan·Mamoru Noda
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Nov 8, 2020·International Journal of Environmental Research and Public Health·Beatrice CasiniGaetano Privitera
Mar 9, 2018·Experimental Parasitology·Nathan H Vande BurgtAnnetta Zintl
Jul 23, 2021·The Science of the Total Environment·Wei HongHongping Wei
Aug 29, 2021·Viruses·Louisa F Ludwig-BegallEtienne Thiry
Jan 25, 2020·Environmental Science & Technology·Nicole RockeyKrista R Wigginton

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