Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland.

Menton, John F; Kearney, Karen; Morgan, John G

dc.contributor.author	Menton, John F
dc.contributor.author	Kearney, Karen
dc.contributor.author	Morgan, John G
dc.date.accessioned	2010-04-06T10:17:17Z
dc.date.available	2010-04-06T10:17:17Z
dc.date.issued	2007
dc.identifier.citation	Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland. 2007, 4:86 Virol. J.	en
dc.identifier.issn	1743-422X
dc.identifier.pmid	17822552
dc.identifier.doi	10.1186/1743-422X-4-86
dc.identifier.uri	http://hdl.handle.net/10147/95628
dc.description.abstract	BACKGROUND: Noroviruses are the most common cause of non-bacterial gastroenteritis. Improved detection methods have seen a large increase in the number of human NoV genotypes in the last ten years. The objective of this study was to develop a fast method to detect, quantify and genotype positive NoV samples from Irish hospitals. RESULTS: A real-time RT-PCR assay and a Reverse Line Blot Hybridisation assay were developed based on the ORF1-ORF2 region. The sensitivity and reactivity of the two assays used was validated using a reference stool panel containing 14 NoV genotypes. The assays were then used to investigate two outbreaks of gastroenteritis in two Irish hospitals. 56 samples were screened for NoV using a real-time RT-PCR assay and 26 samples were found to be positive. Genotyping of these positive samples found that all positives belonged to the GII/4 variant of NoV. CONCLUSION: The combination of the Real-time assay and the reverse line blot hybridisation assay provided a fast and accurate method to investigate a NoV associated outbreak. It was concluded that the predominant genotype circulating in these Irish hospitals was GII/4 which has been associated with the majority of NoV outbreaks worldwide. The assays developed in this study are useful tools for investigating NoV infection.
dc.language.iso	en	en
dc.subject.mesh	Caliciviridae Infections
dc.subject.mesh	Disease Outbreaks
dc.subject.mesh	Gastroenteritis
dc.subject.mesh	Genotype
dc.subject.mesh	Humans
dc.subject.mesh	Ireland
dc.subject.mesh	Norovirus
dc.subject.mesh	Nucleic Acid Hybridization
dc.subject.mesh	Reverse Transcriptase Polymerase Chain Reaction
dc.subject.mesh	Sensitivity and Specificity
dc.title	Development of a real-time RT-PCR and Reverse Line probe Hybridisation assay for the routine detection and genotyping of Noroviruses in Ireland.	en
dc.contributor.department	Lab 439, Food Science Building, Department of Microbiology, University College Cork, Cork, Republic of Ireland. j.menton@ucc.ie	en
dc.identifier.journal	Virology journal	en
refterms.dateFOA	2018-09-03T10:43:06Z
html.description.abstract	BACKGROUND: Noroviruses are the most common cause of non-bacterial gastroenteritis. Improved detection methods have seen a large increase in the number of human NoV genotypes in the last ten years. The objective of this study was to develop a fast method to detect, quantify and genotype positive NoV samples from Irish hospitals. RESULTS: A real-time RT-PCR assay and a Reverse Line Blot Hybridisation assay were developed based on the ORF1-ORF2 region. The sensitivity and reactivity of the two assays used was validated using a reference stool panel containing 14 NoV genotypes. The assays were then used to investigate two outbreaks of gastroenteritis in two Irish hospitals. 56 samples were screened for NoV using a real-time RT-PCR assay and 26 samples were found to be positive. Genotyping of these positive samples found that all positives belonged to the GII/4 variant of NoV. CONCLUSION: The combination of the Real-time assay and the reverse line blot hybridisation assay provided a fast and accurate method to investigate a NoV associated outbreak. It was concluded that the predominant genotype circulating in these Irish hospitals was GII/4 which has been associated with the majority of NoV outbreaks worldwide. The assays developed in this study are useful tools for investigating NoV infection.