Reference : Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay...
Scientific journals : Article
Life sciences : Food science
http://hdl.handle.net/2268/6872
Detection and quantification of human and bovine novoviruses by a TaqMan RT-PCR assay with a control for inhibition.
English
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 mailto [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 mailto [Université de Liège - ULg > Département de sciences des denrées alimentaires > Microbiologie des denrées alimentaires >]
Daube, Georges mailto [Université de Liège - ULg > Département des maladies infectieuses et parasitaires > Virologie, épidémiologie et pathologie des maladies virales >]
Thiry, Etienne mailto [> >]
2008
Molecular and Cellular Probes
22
215-222
Yes (verified by ORBi)
International
0890-8508
[en] Norovirus ; real-time PCR ; diagnosis
[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.
Service public fédéral Santé publique, Sécurité de la Chaîne alimentaire et Environnement ; Service public fédéral de Programmation Politique scientifique
RF6/85 et SD/AF/01
Researchers ; Students
http://hdl.handle.net/2268/6872

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