ORBi: Scipioni Alexandra - Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay with a control for inhibition.

Reference : Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay wi...


	Document type :	Scientific journals : Article
	Discipline(s) :	Life sciences : Food science
	To cite this reference:	http://hdl.handle.net/2268/6872

Title :	Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay with a control for inhibition.
Language :	English
Author, co-author :	Scipioni, Alexandra [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales >]
	Bourgot, Isabelle [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales >]
	Mauroy, Axel [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales >]
	Ziant, Dominique [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Epidémiologie et analyse des risques appl. aux sc. vétér. >]
	Saegerman, Claude [Université de Liège - ULg > Département de sciences des denrées alimentaires > Microbiologie des denrées alimentaires >]
	Daube, Georges [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales >]
	Thiry, Etienne [> >]
Publication date :	2008
Journal title :	Molecular and Cellular Probes
Volume :	22
Pages :	215-222
Audience :	International
ISSN :	0890-8508
Keywords :	[en] Norovirus ; real-time PCR ; diagnosis
Abstract :	[en] Noroviruses are single-stranded RNA viruses belonging to the family Caliciviridae. They are a major cause of epidemic and sporadic gastroenteritis in humans and calves. Reverse transcription-polymerase chain reaction (RT-PCR) has become the ‘‘gold standard’’ for detection of noroviruses in faecal and environmental samples. However, false negative results due to co-concentration of RT-PCR inhibitors are a continuous concern. A TaqMan real-time RT-PCR assay making use of a foreign internal RNA control and a RNA standard was developed. Very interestingly, this method is capable of detecting human noroviruses belonging to genogroups I and II, and bovine noroviruses belonging to genogroup III. Inhibitors were removed efficiently by 1/10 dilution of the sample or addition of bovine serum albumin to the RT-PCR mix. This assay was validated with human and bovine stool samples previously tested for norovirus by conventional RT-PCR. The ability to detect norovirus in stool samples that were negative by conventional RT-PCR assay demonstrate the higher sensitivity of the TaqMan assay compared to the conventional RT-PCR assay. This real-time RT-PCR assay allows the detection of both human and bovine noroviruses, avoids false negative results and is able to quantify the level of norovirus contamination.
Funders :	Service public fédéral Santé publique, Sécurité de la Chaîne alimentaire et Environnement ; Service public fédéral de Programmation Politique scientifique
Name of the research project :	RF6/85 et SD/AF/01
Target :	Researchers ; Students
Permalink :	http://hdl.handle.net/2268/6872

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