PMID: 11337037May 5, 2001Paper

Improved method for the recovery of hepatitis A virus from oysters

Journal of Virological Methods
J L MullendoreY Carol Shieh

Abstract

Hepatitis A is one of the major infectious diseases epidemiologically associated with worldwide shellfish consumption. Molecular detection using polymerase chain reaction (PCR) to detect hepatitis A virus (HAV) in contaminated shellfish can be hindered by low virus recoveries during the concentration process and by natural PCR inhibitors in shellfish. This study evaluated and modified two major steps of a processing procedure for virus concentration from oysters: acid adsorption-elution and solvent extraction. With the addition of second and third elutions, the acid adsorption-elution step doubled the recovery to 46% of HAV seeded initially. Extraction with chloroform or chloroform-butanol resulted in lower HAV detection limits by reverse transcription-PCR (RT-PCR)-oligoprobing than extraction with the fluorocarbon, Freon. These results led to the following modified procedure: HAV was acid adsorbed at pH 4.8, eluted first with 0.05 M glycine, second with 0.5 M threonine, PEG-precipitated twice, chloroform-extracted twice, RNA-extracted, and RT-PCR (single round) amplified. Using the modified procedure, HAV was detected by RT-PCR in all trials with a seeding density of > or = 1 plaque forming unit (PFU)/g of oyster, and in which...Continue Reading

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Citations

Feb 23, 2013·Food and Environmental Virology·M YavarmaneshM R Nassiri
Mar 5, 2004·International Journal of Food Microbiology·Ester SuñénCarmen Zigorraga
Oct 7, 2005·Epidemiology and Infection·H GuisJ F Viel
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Mar 2, 2010·Applied and Environmental Microbiology·Angelo DePaolaKhamphet Nabe
Apr 5, 2003·Applied and Environmental Microbiology·Christian BeuretJakob Schluep
Apr 29, 2014·International Journal of Food Microbiology·Y Carol ShiehDonald W Schaffner
Dec 6, 2011·International Journal of Food Microbiology·Ambroos StalsMieke Uyttendaele
Oct 13, 2007·Journal of Virological Methods·Efstathia PapafragkouLee-Ann Jaykus
Sep 29, 2007·International Journal of Food Microbiology·Nerea CasasIñigo Martínez de Marañón
Dec 22, 2006·International Journal of Food Microbiology·Anna Charlotte SchultzBirgit Nørrung
Oct 5, 2015·International Journal of Food Microbiology·Kathrin SzaboReimar Johne
Feb 7, 2006·Water Research·T C M SinceroC R M Barardi
Jul 7, 2010·Journal of Virological Methods·Katrine UhrbrandAnna Charlotte Schultz
Mar 23, 2015·Food and Environmental Virology·Hiroyuki SaitoMamoru Noda
Oct 12, 2005·International Journal of Food Microbiology·Mary SmiddyColin Hill
Aug 30, 2008·Journal of Virological Methods·Hee-Yeon KimGwangPyo Ko
Nov 27, 2018·Journal of Food Protection·Sang-Mu KoDuwoon Kim
Aug 26, 2019·Food and Environmental Virology·Sofia StrubbiaFrançoise S Le Guyader
Sep 26, 2009·Journal of Food Protection·Sooryun CheongSang-Jong Kim
May 5, 2007·Journal of Food Protection·Ana Maria de Roda HusmanSaskia A Rutjes
Apr 4, 2021·Viruses·Yarrow S LindenJohn Scott Meschke

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