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

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    Authors
    Menton, John F
    Kearney, Karen
    Morgan, John G
    Affiliation
    Lab 439, Food Science Building, Department of Microbiology, University College Cork, Cork, Republic of Ireland. j.menton@ucc.ie
    Issue Date
    2007
    MeSH
    Caliciviridae Infections
    Disease Outbreaks
    Gastroenteritis
    Genotype
    Humans
    Ireland
    Norovirus
    Nucleic Acid Hybridization
    Reverse Transcriptase Polymerase Chain Reaction
    Sensitivity and Specificity
    
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    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.
    Journal
    Virology journal
    URI
    http://hdl.handle.net/10147/95628
    DOI
    10.1186/1743-422X-4-86
    PubMed ID
    17822552
    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
    ISSN
    1743-422X
    ae974a485f413a2113503eed53cd6c53
    10.1186/1743-422X-4-86
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