Thumbnail Image
Publication

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
Author
Menton, John F
 Kearney, Karen
 Morgan, John G
Advisors
Editors
Other Contributors
Departments
Lab 439, Food Science Building, Department of Microbiology, University College Cork, Cork, Republic of Ireland. j.menton@ucc.ie
Date
2007
Date Submitted
Keywords
Other Subjects
Subject Mesh
Caliciviridae Infections
Disease Outbreaks
Gastroenteritis
Genotype
Humans
Ireland
Norovirus
Nucleic Acid Hybridization
Reverse Transcriptase Polymerase Chain Reaction
Sensitivity and Specificity
Planned Date
Start Date
Collaborators
Principal Investigators
Files
Thumbnail Image
17822552.pdf
Adobe PDF, 339.85 KB
Alternative Titles
Publisher
URI
https://hdl.handle.net/10147/95628
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.
Language
en
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.
ISSN
1743-422X
eISSN
ISBN
DOI
10.1186/1743-422X-4-86
PMID
17822552
PMCID
Sponsorships
Funding Sources
Funding Amounts
Grant Identifiers
Methodology
Duration
Ethical Approval