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Multiplex PCR to Differentiate Monkeypox Virus Clades.

Christopher T Williams, Alessandra Romero-Ramirez, Adeleye Adesola Semiu

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    This summary is machine-generated.

    A new multiplex quantitative PCR assay effectively differentiates monkeypox virus clades. The assay shows high sensitivity and specificity, particularly for samples with lower cycle thresholds, aiding in accurate monkeypox virus detection.

    Keywords:
    MPXVNigeriaPCRUnited Kingdomdiagnosticsmonkeypox virusmpoxviruseszoonoses

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    Area of Science:

    • Virology
    • Molecular Biology
    • Infectious Diseases

    Background:

    • Monkeypox virus (MPXV) clade differentiation is crucial for epidemiological tracking and public health response.
    • Accurate diagnostic tools are needed to distinguish between MPXV clades, especially during outbreaks.

    Purpose of the Study:

    • To develop and evaluate a multiplex quantitative PCR (qPCR) assay for differentiating MPXV clades.
    • To assess the sensitivity and specificity of the developed assay using clinical samples.

    Main Methods:

    • Design of a multiplex quantitative PCR assay targeting specific regions of the MPXV genome.
    • Testing the assay on clinical samples from the United Kingdom and Nigeria.
    • Analysis of assay performance, including sensitivity and specificity, with a focus on cycle threshold values.

    Main Results:

    • The multiplex qPCR assay demonstrated good performance in differentiating MPXV clades.
    • Overall sensitivity was 78% and specificity was 94% for clinical samples.
    • For samples with cycle thresholds <35, sensitivity increased to 98% and specificity remained at 94%.

    Conclusions:

    • The developed multiplex qPCR assay is a valuable tool for accurate MPXV clade differentiation.
    • The assay's high performance, especially with low cycle threshold samples, supports its clinical utility.
    • This assay can aid in real-time epidemiological surveillance and outbreak management of monkeypox.