Optimisation of a single-primer sequence-independent amplification (SP-SIA) assay: Detection of previously undetectable norovirus strains associated with outbreaks of gastroenteritis

S. Braham, M. Iturriza-Gómara, J. Gray*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    5 Citations (Scopus)

    Abstract

    The introduction of molecular diagnostic methods for investigation of gastroenteritis has significantly reduced the diagnostic gap. However, approximately 25% of cases of gastroenteritis remain undiagnosed even after screening for bacteria, parasites and viruses using the most sensitive PCR and RT-PCR methods available. In recent years, it has become apparent that viruses are responsible for the majority of outbreaks of gastroenteritis. In this study, a panel of samples from outbreaks of gastroenteritis for which no aetiological agent had been identified was selected for investigation by random amplification molecular methods. An algorithm for virus purification and concentration was developed followed by a single-primer sequence-independent amplification method. These methods resulted in the identification of viruses in 5 out of 51 previously negative outbreaks. Noroviruses undetectable using two available broadly reactive diagnostic methods were detected in 4 of these outbreaks.

    Original languageEnglish
    Pages (from-to)30-34
    Number of pages5
    JournalJournal of Virological Methods
    Volume158
    Issue number1-2
    DOIs
    Publication statusPublished - Jun 2009

    Bibliographical note

    Funding Information:
    This work was supported through an EUFW6 grant (EVENT: SP22-CT-2004-502571). We would like to thank Prof. Polly Roy, LSHTM, for helpful advice throughout the project.

    Copyright:
    Copyright 2009 Elsevier B.V., All rights reserved.

    Keywords

    • Gastroenteritis
    • Norovirus
    • Outbreaks
    • Sequence-independent amplification (SIA)

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