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Sensitive Multiplex Real-time RT-qPCR Assay for the Detection of Filoviruses

Vladimir G Dedkov, N'Faly Magassouba, Marina V Safonova

    Health Security
    |January 20, 2018
    PubMed

    Insights

    A new real-time RT-qPCR assay offers rapid and sensitive detection of filoviruses, crucial for managing emergent diseases globally. This diagnostic tool aids clinical and epidemiological investigations of filovirus fever diseases.

    Area of Science:

    • Virology
    • Molecular Diagnostics
    • Public Health

    Background:

    • Filoviruses cause emergent diseases with high mortality rates, posing a global health risk due to increased interconnectedness.
    • Effective management of filovirus risk requires reliable diagnostic tools, with real-time reverse transcription PCR (RT-qPCR) being the most suitable method.

    Purpose of the Study:

    • To develop and evaluate a novel real-time RT-qPCR assay for the rapid and sensitive detection of filoviruses.
    • To enhance existing diagnostic capabilities for filovirus fever diseases.

    Main Methods:

    • Development of two sets of real-time RT-qPCR assays (FiloA-Fl and FiloB-Fl) for filovirus detection.
    • Evaluation of the assays using armored RNA phage particles (ARs) as positive controls.
    • Determination of the limit of detection for each assay set.

    Main Results:

    • The FiloA-Fl assay demonstrated a limit of detection of 5x10^2 copies/ml of AR-positive control.
    • The FiloB-Fl assay showed a limit of detection of 5x10^3 copies/ml of AR-positive control.
    • The developed assay proved to be rapid, sensitive, and specific for filovirus detection.

    Conclusions:

    • The new real-time RT-qPCR assay is a valuable tool for rapid and sensitive filovirus detection.
    • The assay's high specificity and sensitivity support its use in clinical and epidemiological investigations of filovirus diseases worldwide.
    Keywords:
    DiagnosticsEbola virusFilovirusesMarburg virusRT-qPCR

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