Development of a method for direct extraction of viral RNA from bivalve molluscs

Letters in Applied Microbiology
H Quang LeD De Medici

Abstract

The detection of foodborne viruses in bivalve molluscs is a challenging procedure in relation to low virus concentration and to the presence of significant RT-PCR inhibitors. The aim of this study was the development of an efficient direct extraction method for foodborne viral RNA from bivalve molluscs. Using Mengovirus as a surrogate for foodborne viruses, five extraction methods based on RNA release by Trizol were compared on clams and oysters. A procedure consisting of Trizol, PureLink RNA Mini Kit, followed by Cetyltrimethylammonium bromide (CTAB) treatment and LiCl precipitation was found to provide RNA with the highest extraction efficiency and negligible inhibitory effect on real-time RT-PCR. This procedure was further compared to standard extraction method (ISO 15216) using clam, mussel and oyster samples spiked with Hepatitis A virus, Norovirus (NoV) GI and GII as well as bivalve samples naturally contaminated with NoV GI or GII. Results clearly demonstrated that the developed method provided, on average, a recovery 4·3 times higher than the standard reference protocol as well as good repeatability. A direct extraction procedure was developed to recover viral RNA from shellfish with improved efficiency in comparison to...Continue Reading

References

Nov 1, 1991·The Journal of Infectious Diseases·M L HallidayS L Hu
Jun 1, 1980·European Journal of Biochemistry·C Auffray, F Rougeon
Aug 1, 1994·Applied and Environmental Microbiology·D N LeesW J Doré
Feb 1, 1993·Applied and Environmental Microbiology·R L AtmarM K Estes
Mar 11, 2005·Emerging Infectious Diseases·Goutam K AdakSarah J O'Brien
Feb 28, 2006·International Journal of Food Microbiology·Ingeborg L A BoxmanMarion Koopmans
Jul 22, 2008·International Journal of Food Microbiology·Heidi Lange ComelliMette Myrmel
Jun 3, 2010·Cold Spring Harbor Protocols·Donald C RioTimothy W Nilsen
Jul 7, 2010·Journal of Virological Methods·Katrine UhrbrandAnna Charlotte Schultz
Jan 5, 2011·Emerging Infectious Diseases·Elaine ScallanPatricia M Griffin
Feb 1, 2012·Journal of Food Protection·James A LowtherDavid N Lees
May 1, 2012·International Journal of Food Microbiology·Arie H HavelaarWilfrid van Pelt
Jun 12, 2012·Journal of Applied Microbiology·E SuffrediniL Croci
Jul 2, 2014·The Lancet Infectious Diseases·Sharia M AhmedBenjamin A Lopman
Jul 13, 2014·International Journal of Food Microbiology·J S KrogA C Schultz
Dec 3, 2014·International Journal of Food Microbiology·David PoloJesús L Romalde
Dec 15, 2015·Applied and Environmental Microbiology·Miguel F VarelaJesús L Romalde
Apr 30, 2017·Food and Environmental Virology·Elisabetta SuffrediniDario De Medici
May 4, 2018·Food and Environmental Virology·J A LowtherD N Lees
Apr 1, 2016·Current Opinion in Food Science·Albert BoschSusana Guix

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Citations

Jun 25, 2019·Transboundary and Emerging Diseases·Ravo M RazafimahefaEtienne Thiry
Jul 26, 2019·Food and Environmental Virology·Thamapan TunyakittaveewardLeera Kittigul
Mar 12, 2021·Frontiers in Microbiology·Amy H FitzpatrickPaul Cotter
Jun 30, 2021·Critical Reviews in Food Science and Nutrition·Min YangDeqing Zhou

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