{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T11:43:46Z","timestamp":1788263026803,"version":"build-2803163510"},"reference-count":106,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Technology Innovation Agency","award":["B1B0741B-9691-4333-B77E-3B8D42A7B5FF"],"award-info":[{"award-number":["B1B0741B-9691-4333-B77E-3B8D42A7B5FF"]}]},{"name":"Technology Innovation Agency","award":["C6M0248"],"award-info":[{"award-number":["C6M0248"]}]},{"name":"Technology Innovation Agency","award":["C1M0224"],"award-info":[{"award-number":["C1M0224"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: Monkeypox (MPX), caused by the Monkeypox virus (MPOX) of the Orthopoxvirus genus, has re-emerged as a significant global health threat. Once confined to Central and West Africa, the 2022\u20132025 multi-country outbreaks, predominantly caused by Clade IIb, demonstrated sustained human-to-human transmission and global spread. Objective: This review summarizes current knowledge on MPX virology, epidemiology, clinical presentation, and diagnostic technologies, with a focus on innovations supporting rapid and field-deployable detection in resource-limited settings. Methods: The recent literature (2019\u20132025), including peer-reviewed studies, WHO and Africa CDC reports, and clinical guidelines, was critically reviewed. Data were synthesized to outline key developments in diagnostic methodologies and surveillance approaches. Results: MPX comprises two genetic clades: Clade I (Congo Basin) and Clade II (West African), which differ in virulence and transmission. Clade IIb is associated with sexual and close-contact transmission during recent outbreaks. Clinical manifestations have shifted from classic disseminated rash to localized anogenital lesions and atypical or subclinical infections. RT-PCR remains the diagnostic gold standard, while emerging assays such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and CRISPR\/Cas-based platforms show promise for rapid point-of-care (POC) testing. Complementary serological tools, including ELISA and lateral flow assays, enhance surveillance and immune profiling. Conclusions: The resurgence of MPX highlights the urgent need for accessible, sensitive, and specific diagnostic platforms to strengthen surveillance and outbreak control, especially in endemic and resource-constrained regions.<\/jats:p>","DOI":"10.3390\/diagnostics15232991","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T10:46:18Z","timestamp":1764067578000},"page":"2991","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Diagnostic Advances and Public Health Challenges for Monkeypox Virus: Clade-Specific Insight and the Urgent Need for Rapid Testing in Africa"],"prefix":"10.3390","volume":"15","author":[{"given":"Caroline N.","family":"Sambo","sequence":"first","affiliation":[{"name":"Human Diagnostic Platform, Chemicals Cluster, Council for Scientific and Industrial Research, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8121-5617","authenticated-orcid":false,"given":"Amanda","family":"Skepu","sequence":"additional","affiliation":[{"name":"Human Diagnostic Platform, Chemicals Cluster, Council for Scientific and Industrial Research, Pretoria 0001, South Africa"},{"name":"Department of Life and Consumer Sciences, College of Agriculture and Environmental Sciences, University of South Africa, Private Bag X6, Florida, Johannesburg 1710, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nolwandle P.","family":"Nxumalo","sequence":"additional","affiliation":[{"name":"Food Safety Group, Advanced Agriculture Food Cluster, Council for Scientific and Industrial Research, Pretoria 0001, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ketlareng L.","family":"Polori","sequence":"additional","affiliation":[{"name":"Faculty of Health Science, Division of Public Health, University of Free State, Bloemfontein Campus, 205 Nelson Mandela Drive, Park West, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Harapan, H., Ophinni, Y., Megawati, D., Frediansyah, A., Mamada, S.S., Salampe, M., Bin Emran, T., Winardi, W., Fathima, R., and Sirinam, S. 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