An accurate and rapid Real-time PCR approach for human Monkeypox virus diagnosis

BackgroundHuman monkeypox (MPX) is an emerging zoonotic disease caused by monkeypox virus (MPXV), which is endemic to Western and Central Africa. Despite being identified in captive monkeys in 1958, and in humans from the Democratic Republic of Congo (DRC) in 1970, a MPXV diagnostic test is not routinely implemented in diagnostic laboratories. Currently, a multi-country outbreak of MPXV has been ongoing in several European and American countries since May 2022. MethodsTo develop this real-time PCR assay, five MPXV positive DNA samples were tested using MPXV-specific primers. The qPCR amplification was conducted on two different qPCR devices. We also tested samples, collected in May 2022, from pox-like lesions of MPXV-suspected patients in Belgium. ResultsThe performance of our real-time PCR was proved with five MPXV positive DNA from DRC. In addition, five out of ten clinical samples, from the current MPXV-outbreak, tested positive with Ct values ranging from 20.6 to 34.9. ConclusionsIn this study, we present an accurate and rapid real-time PCR approach, based on conserved regions of both MPXV clades, that can be used to test clinical samples from MPXV-suspected persons. Our real-time qPCR assay can be used for the 2022 multi-country outbreak of human MPXV. SummaryAn accurate and fast real-time PCR assay is indispensable to diagnose cases of monkeypox during the 2022 multi-country outbreak.