Evaluation of a single-tube fluorogenic RT-PCR assay for detection of bovine respiratory syncytial virus in clinical samples

Mikhayil Hakhverdyan, Sara Hägglund, Lars Erik Larsen, Sándor Belák

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Bovine respiratory syncytial virus (BRSV) causes severe disease in naive cattle of all ages and is a common pathogen in the respiratory disease complex of calves. Simplified methods for rapid BRSV diagnosis would encourage sampling during outbreaks and would consequently lead to an extended understanding of the virus. In this study, a BRSV fluorogenic reverse transcription PCR (fRT-PCR) assay, based on TaqMan principle, was developed and evaluated on a large number of clinical samples, representing various cases of natural and experimental BRSV infections. By using a single-step closed-tube format, the turn-around time was shortened drastically and results were obtained with minimal risk for cross-contamination. According to comparative analyses, the detection limit of the fRT-PCR was on the same level as that of a nested PCR and the sensitivity relatively higher than that of a conventional PCR, antigen ELISA (Ag-ELISA) and virus isolation (VI). Interspersed negative control samples, samples from healthy animals and eight symptomatically or genetically related viruses were all negative, confirming a high specificity of the assay. Taken together, the data indicated that the fRT-PCR assay can be applied to routine virus detection in clinical specimens and provides a rapid and valuable too] in BRSV research.
    Original languageEnglish
    JournalJournal of Virological Methods
    Volume123
    Issue number2
    Pages (from-to)195-202
    ISSN0166-0934
    DOIs
    Publication statusPublished - 2005

    Keywords

    • detection
    • TaqMan
    • real-time
    • BRSV
    • fluorogenic RT-PCR

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