ISSN: 1859 - 2872
Optimize real-time PCR reaction to detect monkeypox virus in human
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Abstract
Objective: The study aims to the optimization of a highly sensitive and specific real-time PCR reaction for the detection of the monkeypox virus. Subject and method: This real-time PCR-based method was employed for the amplification of target genes, utilizing relevant negative controls, and a monkeypox virus plasmid control sourced from the CDC. Detection limits and sensitivity were determined through a range of spiking assays covering a concentration range from 106 to 100copies/µL. Result: Our data demonstrated that the Limit of Detection (LOD) of our method for detecting monkeypox virus DNA is 250 copies per milliliter (mL). The assay showed a high level of specificity at 100% and a sensitivity of 95%. Conclusion: In conclusion, we have successfully established a highly sensitive and specific real-time PCR assay for the detection of monkeypox virus using a positive standard DNA plasmid template equivalent to the CDC protocol.
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