Performance of pre-RT-qPCR treatments to discriminate infectious human rotaviruses and noroviruses from heat-inactivated viruses: applications of PMA/PMAxx, benzonase and RNase

Journal of Applied Microbiology
S OristoL Maunula

Abstract

Detection/Quantification of RNA viruses is mostly done by reverse-transcriptase (RT)-(q)PCR, but it does not distinguish between infectious and noninfectious viruses. Our aim was to test, how different pretreatments before RT-qPCR could eliminate positivity originated from external nucleic acids or genomes of damaged particles. Heat-inactivated (80°C for 10 min) rotavirus Wa strain and faecal samples containing rotavirus or norovirus were treated with PMA/PMAxx, benzonase or crude extract RNase prior to RT-qPCR. PMA/PMAxx pretreatments were not consistently efficient for RV, although they seemed to work to some extent for heat-inactivated norovirus. Benzonase and RNase provided consistently 2·2-2·8 log10reductions in the titre of faecal rotavirus. All pretreatments need to be further validated for each virus separately, taking into account sample matrix and inactivation conditions. Although none of the pretreatments could completely render inactivated viruses undetectable, RNase worked most consistently for both rota- and norovirus. This study sheds light on capacity of the most common pre-RT-qPCR treatments to eliminate damaged, noninfectious rotaviruses and noroviruses after thermal treatment. To our knowledge, this is the fi...Continue Reading

References

Jun 29, 2002·Journal of Virological Methods·Suphachai Nuanualsuwan, Dean O Cliver
Mar 5, 2005·Applied and Environmental Microbiology·W J Lodder, A M de Roda Husman
Mar 16, 2007·Journal of Clinical Microbiology·Sanela SvrakaMarion Koopmans
Feb 23, 2008·Water Research·Ana Cecilia EspinosaCarlos F Arias
Sep 4, 2008·Journal of Virological Methods·S Q ZengT Vesikari
Apr 21, 2009·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Tuija KantalaMaija Lappalainen
Nov 4, 2009·Current Protocols in Microbiology·Michelle ArnoldSarah M McDonald
Mar 18, 2010·Food and Environmental Virology·Dean O Cliver
Dec 16, 2010·The EMBO Journal·Ethan C SettembreStephen C Harrison
Aug 17, 2011·Journal of Virological Methods·Dan LiMieke Uyttendaele
Feb 16, 2013·Food and Environmental Virology·Gloria SánchezRosa Aznar
Oct 15, 2013·Journal of Applied Microbiology·P ElizaquívelG Sánchez
Apr 11, 2014·Human Vaccines & Immunotherapeutics·Ming Tan, Xi Jiang
Jul 23, 2014·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Jacqueline E Tate, Umesh D Parashar
Mar 10, 2015·International Journal of Hygiene and Environmental Health·Mats LeifelsIbrahim Ahmed Hamza
Mar 23, 2015·Journal of Virological Methods·Mohammad R KarimSandhya U Parshionikar
Apr 10, 2016·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Jacqueline E TateUNKNOWN World Health Organization–Coordinated Global Rotavirus Surveillance Network
Jul 1, 2014·Comprehensive Reviews in Food Science and Food Safety·Siele CeuppensJohn Donaghy

❮ Previous
Next ❯

Citations

May 21, 2021·Frontiers in Microbiology·Elisenda BallestéCristina García-Aljaro
Mar 14, 2021·Molecular Ecology Resources·Patricia GilSerafin Gutierrez
Jan 25, 2020·Environmental Science & Technology·Nicole RockeyKrista R Wigginton

❮ Previous
Next ❯

Related Concepts

Related Feeds

Anxiety Disorders

Discover the latest research on anxiety disorders including agoraphobia, panic disorder, obsessive-compulsive disorder, and post-traumatic stress disorder here.