A critical assessment of two real-time PCR assays targeting the (SSU) rRNA and gdh genes for the molecular identification of Giardia intestinalis in a clinical laboratory

S. Boadi, S. D. Polley, Sally Kilburn, Graham A. Mills, P. L. Chiodini

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    Abstract

    Introduction
    Giardiasis is an intestinal diarrhoeal illness caused by the flagellate protozoan parasite Giardia intestinalis. Molecular techniques for the identification of G. intestinalis have generally been shown to offer a better detection rate of the parasite than the traditional faecal concentration and microscopy techniques.

    Aim
    The aim of this study was to critically assess the performance of a commercial and a published real-time PCR assay for their potential use as frontline tests for the diagnosis of giardiasis.

    Methods
    A composite reference standard of enzyme immunoassay and rapid membrane test was used in a diagnostic accuracy study to assess the performance of Primerdesign's, and Verweij et al G. intestinalis real-time PCR assays, comparing them with the traditional ova, cysts and parasite microscopy test (OCP-M).

    Results
    The Verweij real-time PCR used primers for the (SSU) rRNA gene, and produced a diagnostic sensitivity of 93.4% (95% CI 88.30% to 98.50%) and an efficiency of 100%. Primerdesign's real-time PCR used primers for the glutamate dehydrogenase gene and produced a diagnostic sensitivity of 61.5% (95% CI 51.50% to 71.50%) and an efficiency of 203%. The OCP-M sensitivity was 83.5% (95% CI 75.87% to 91.13%).

    Conclusions
    The Verweij real-time PCR was robust and the most sensitive assay suited for use as a first-line diagnostic test for giardiasis.
    Original languageEnglish
    Pages (from-to)811-816
    JournalJournal of Clinical Pathology
    Volume67
    Issue number9
    Early online date11 Jun 2014
    DOIs
    Publication statusPublished - 2014

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