Monkeypox disease develops from the monkeypox virus, which contains double-stranded DNA and belongs to the orthopoxvirus genus found under the family Poxviridae, subfamily Chordopoxvirinae. The WHO director declared the multiregional monkeypox outbreak an international public health emergency. This review aims to summarise the pathogenesis, virology, epidemiology, clinical features, diagnostics, the available therapeutic and prophylactic measures, as well as future perspectives, research gaps, and global preparedness for monkeypox. Monkeypox virus has an architectural resemblance to the variola virus but has its own genetic and pathogenic characteristics. Monkeypox virus has evolved globally to spread from specific African regions to cause human-to-human transmission in other worldwide territories. Standard clinical signs, including fever and headache pain along with muscle pain, swollen lymph nodes, and rashes in specific body regions, manifest as the first step of diagnosis. Traditional polymerase chain reaction together with real-time polymerase chain reaction (RT-PCR) stands as the main diagnostic tool that healthcare providers use with imaging studies for complication assessment. Some antiviral drugs, including tecovirimat, cidofovir, and brincidofovir, are FDA-approved for the treatment of smallpox and are used for mpox under the Expanded Access Investigational New Drug (EA-IND) protocol or other authorised investigational/off-label settings for eligible patients. JYNNEOS (IMVANEX) is approved by the FDA and EMA for the prevention of both smallpox and mpox, whereas ACAM2000 is approved for smallpox and may be used for mpox in specific off-label or public health settings. The primary preventive strategy involves community-oriented preventive initiatives for infectious diseases, including future studies regarding gaps in vaccinations, the dynamics of transmission, and enhancing the preparedness of global health systems. The comprehensive review reveals Mpox disease history and current knowledge through a systematic evaluation of disease source, virology, disease transmission and clinical presentation, detection, management and prevention approaches and future plans to stop this infection.

The continuing evolution of monkeypox virus (MPXV) clades capable of sustained human-to-human transmission poses an escalating global health risk, yet no effective therapy exists for poxvirus infection. Here, we investigate a panel of human monoclonal antibodies (mAbs), derived from an antibody library, which targets A35 and B6 antigens located on the extracellular enveloped virion (EV). Four high-affinity mAbs target the central groove of the A35 dimer, whereas one targets a previously unidentified epitope on the A35 protruding loop. Three other mAbs recognize distinct, previously unmapped epitopes on the multidomain B6 antigen. Those epitopes are conserved across most orthopoxviruses. A combination of an anti-A35 and an anti-B6 antibody provides additive and complete protection from both morbidity and mortality in a lethal VACV mouse model, challenging the previous assumption that targeting both the EV and mature virion (MV) forms is required for full protection. These studies uncover critical information for next-generation therapeutics and vaccines against emerging MPXV as well as other Orthopoxvirus threats.

Monkeypox virus (MPXV) continues to emerge as a global health threat. While genetically diverse, its genomic structure and evolution remain undercharacterized. This study analyzed 157 non-redundant MPXV genomes (subtypes Ia, Ib, IIa, IIb), identifying 183 core and 222 non-core genes via pan-genome analysis-indicative of an open pan-genome. Over half (55.8%) of MPXV genes show gene number variation. Non-core genes exhibit a higher ratio of insertions/deletions compared to core genes. Additionally, selection pressure analysis indicated that non-core genes are more prone to positive selection. Furthermore, interaction analysis revealed that core viral proteins target immune-related human pathways, while non-core proteins engage with viral infection-associated proteins. These results deepen our understanding of MPXV evolution and inform future research directions.

Real-time, quantitative dynamic monitoring of multiple biomarkers is urgently needed in clinical practice; however, conventional technologies are limited by cumbersome workflows and time-consuming operational procedures. Surface-enhanced Raman scattering (SERS) offers high sensitivity and molecular fingerprinting capability, but its application in dynamic monitoring is restricted by signal instability caused by signal irreproducibility arising from stochastic hotspot analyte spatial mismatches, and an exclusive reliance on endpoint measurements that precludes kinetic information extraction. Hence, we present an optofluidic SERS chip integrating spatially encoded photonic microsphere lens arrays (SE-PMLAs) with deterministic hotspaces to overcome these limitations. Through precise integration of gold nanoparticles onto microsphere lens surfaces, each functional unit achieves cascaded photonic-plasmonic coupling among photonic nanojets, whispering-gallery modes, and localized surface plasmon resonance, with near-field feedback reinforcement that transforms random hotspots into engineered, uniform hotspaces, delivering an average enhancement factor of 2.8 × 1010 and signal uniformity with a relative standard deviation below 5%. Using spatially encoded sensing, we demonstrate single-sample, synchronous high-throughput detection of influenza A/B viruses, adenovirus, and monkeypox virus, achieving limits of detection of 0.53 pg/mL, 128.85 copies/mL, 199.56 copies/mL, and 2.7 pg/mL, respectively. Crucially, the chip's ultrastable signal output enables a kinetic quantification model based on the initial binding rate, revealed a linear correlation between analyte concentration and its initial binding rate (i.e., slope). This approach achieves rapid, high-precision dynamic monitoring within 60 to 120s while expanding the linear detection range by two orders of magnitude. Ultimately, this chip delivers a robust, minute-level point-of-care solution for real-time pathogen screening and critical disease diagnostics.

Monkeypox virus (MPXV) is an ongoing global public health concern. Historically associated with zoonotic infection, in 2022, a major epidemiological shift occurred when sustained transmission was identified across multiple countries that previously had not had endemic circulation of the virus. While mpox typically presents with characteristic clinical features, infection can also lead to a spectrum of less common manifestations involving multiple organ systems. The extent of viral dissemination and disease severity appear to be modulated by host factors such as immune status, vaccination history, co-infections, and potentially the viral clade. Although complications associated with mpox are infrequent, they can lead to significant morbidity and mortality. Moreover, the underlying pathogenesis and to what extent the viral clade may be playing a role remain incompletely understood. This review summarizes the less common clinical complications of mpox, explores the underlying mechanisms of viral pathogenesis, and discusses what insights can be gained from cell culture and animal models. Additionally, we highlight knowledge gaps that can guide future research priorities.

Background/Objectives: Recent outbreaks of the mpox virus (MPVX) have highlighted the threat posed by the Orthopoxvirus (OPXV) genus, which includes variola, the causative agent of smallpox. Since routine Vaccinia virus (VACV) vaccination campaigns terminated, the majority of the global population currently lacks immunity against OPXVs. Methods: Using enzyme-linked immunosorbent assays (ELISAs), this study evaluated antibody titers against both VACV proteins and their MPVX orthologs across various age groups. Cross-reactivity was analyzed using 220 serum samples collected from Albanian individuals between November 2023 and January 2024, arranged into five age cohorts. We measured plasma antibody levels against whole VACV, as well as specific surface antigens: L1, B5, A33, and A27 for VACV, and M1, H3, A35, and A29 for MPXV. Results: The data reveal a long-term, gender-independent immunological memory targeting both whole-virus particles and individual viral proteins. Conclusions: Notably, the oldest cohort exhibited higher antibody titers against specific single viral proteins, whereas the younger vaccinated groups showed stronger activity against the whole virus. Additionally, antibodies raised via VACV vaccination recognize MPXV ortholog proteins equally well. This cross-reactivity highlights the broad protective potential of VACV immunization across the entire Orthopoxvirus genus.

Mpox is a zoonotic disease caused by the monkeypox virus, a double-stranded DNA virus belonging to the genus Orthopoxvirus. The World Health Organization (WHO) declared mpox a Public Health Emergency of International Concern (PHEIC) in July 2022 amid the global clade IIb outbreak and again in August 2024 following an increase in cases in the Democratic Republic of the Congo associated with clades Ia and Ib. Despite increased awareness, diagnosing mpox remains challenging because its clinical manifestations often overlap with those of other infectious and non-infectious conditions. This diagnostic challenge occurs across geographic settings but may be further complicated in tropical regions by the circulation of locally prevalent infections, including arboviral diseases such as dengue and chikungunya. Here, we describe a series of nine patients initially evaluated for suspected mpox, in whom alternative diagnoses were subsequently established.

Single-reaction multiplex nucleic acid testing (NATs) must both co-amplify multiple targets and distinguish the resulting amplicons within the product mixture. Most conventional assays achieve this distinction using target-specific reporters or other external encoding elements. Here, we introduce a primer-programmed multiplexing strategy in which primer design generates target-associated amplicons with predefined lengths that are read electrically with nanopores. In a proof-of-concept implementation, one-pot recombinase polymerase amplification (RPA) generated four amplicons with predefined product lengths from Monkeypox (Mpox), Cowpox, Vaccinia, and human RNase P. The resulting amplicons were analyzed with glass nanopores, and individual translocation events were classified by a deep neural network (DNN) based on a six-dimensional feature set derived from nanopore recordings. Event-level decoding achieved >95% accuracy, >94.7% sensitivity, and >98.3% specificity, and aggregation of classified events yielded sample-level calls with >93.3% sensitivity and >93.9% specificity in blinded samples, with qualitative calls matching polymerase chain reaction (PCR) results across the 9 PCR-benchmarked samples. These results demonstrate a probe-free multiplexing architecture in which target identity is programmed through primer-defined amplicon design and decoded from the resulting multidimensional nanopore response using a DNN, eliminating the need for target-specific reporter chemistry and opening new opportunities for all-electrical multiplex nucleic acid testing based on unmodified amplification products.

Monkeypox is currently the most important orthopoxvirus infection in humans. The increase in the number of cases, combined with its potential social impact and international spread, led the World Health Organization (WHO) to classify it as the most recent Public Health Emergency of International Concern (PHEIC). Although mainly endemic in resource-limited countries, quantitative Polymerase Chain Reaction (qPCR) remains the gold standard for detection but is challenging due to the need for specialized instrumentation. To address this, we propose an instrument-friendly molecular assay based on colorimetric Loop-Mediated Isothermal Amplification (LAMP), performed on a multi-assay polystyrene-toner (PST) centrifugal device. The new analytical device was designed to include chambers for loading the LAMP mixture and samples, interconnected channels for mixing them, and separate chambers for simultaneous LAMP amplification and visual detection of each sample. When evaluated in a panel of real clinical samples, compared to qPCR, the assay based on visual detection demonstrated 100.00% specificity, 91.67% sensitivity, and 95.83% accuracy, which increased to 100% accuracy when digital image analysis was employed, with a total analysis time of approximately 60 min. The relative simplicity of the assay may facilitate its implementation in resource-limited settings and contribute to expanding access to molecular detection of monkeypox.

Real-time and highly sensitive monitoring of pathogens in environmental and host-derived samples is critical for controlling emerging and acute infectious diseases, yet remains technically challenging. Here, we report a multifunctional nanozyme-based immunochromatographic assay (ICA) that enables rapid, universal, and multiplex detection in different scenarios through visual readouts. We constructed a corona-structured magnetic nanozyme (CSMN) with a triple signal-amplification mechanism involving "magnetic enrichment-synergistic enhancement-multiple catalytic sites," enabling efficient enrichment of trace pathogens in complex matrices and generation of dual visual outputs (colorimetric and catalytically enhanced signals) on ICA. The CSMN-based nanozyme ICA requires only 2 min of catalysis to achieve a ≥92.65-fold increase in sensitivity and a 100-fold expansion of the detection range. Based on this platform, dual-mode detection strategies were developed for SARS-CoV-2 and monkeypox virus. The direct mode enables rapid screening within 12 min, with detection limits as low as 19.4 and 22.5 pg/mL for the two targets, respectively. The enrichment mode enables ultrasensitive detection within 20 min, with detection ranges of 0.0012-1000 ng/mL and 0.0026-1000 ng/mL, respectively, while mitigating interference from the hook effect, making it suitable for detecting low-abundance targets in complex samples. The method demonstrated high accuracy and reliability across environmental and clinical samples, with a detection time approximately one-sixth that of qPCR and ELISA, indicating strong potential for infectious disease surveillance and outbreak control.

Mpox, formerly known as monkeypox, is caused by Monkeypox virus (MPXV) and has been associated since 2022 with exanthematous illness in men who have sex with men (MSM), including HIV-negative individuals without prior MPXV vaccination.[1,2] The clinical course is usually self-limited, with cutaneous and mucosal involvement.[4,10,12] Severe forms, including keratoconjunctivitis and corneal ulcers, are uncommon and have mainly been linked to immunosuppression.[2,3,5,10,12,14] We report a 20-year-old MSM male, HIV-negative, immunocompetent, and using pre-exposure prophylaxis (PrEP). He presented with generalized exanthema, right ocular pain, conjunctival injection, tearing, and periorbital cellulitis.[1,2,5,10,14] Ophthalmologic evaluation confirmed severe keratoconjunctivitis with progressive corneal ulcers.[5-7,10,11,14] MPXV PCR was positive in skin and corneal samples.[1,2,10,14] This case highlights the potential for severe ocular disease in a young, otherwise healthy patient and underscores the need for high clinical suspicion, early ophthalmologic assessment, and timely antiviral therapy.[2,4,10,15].

The second WHO Public Health Emergency of International Concern (PHEIC) for mpox was declared on 14 August 2024 after clade Ib monkeypox virus emerged in the Democratic Republic of the Congo (DRC) and spread rapidly across Africa and internationally. The PHEIC ended on 5 September 2025, and the Africa CDC Public Health Emergency of Continental Security was lifted on 22 January 2026. This study characterised global and Africa-specific mpox epidemiology across the full second PHEIC cycle and immediate post-emergency period, from January 2024 to February 2026. A retrospective surveillance analysis was conducted using the WHO Global Mpox Trends dashboard exported data, supplemented by WHO situation reports S32-S64 and international and national surveillance reports. Analyses included negative binomial segmented regression, travel-linked pathway analysis, exploratory analysis of documented secondary transmission after travel-linked detection, and descriptive statistics. Models used reported case counts with log population offsets; reported case-rate ratios therefore reflect reported case rates rather than true infection incidence. Between January 2024 and February 2026, 82,738 confirmed cases and 306 deaths were reported to WHO globally (case fatality ratio 0.37%). The PHEIC period accounted for 74.0% of cases, peaking at 7,405 cases in May 2025. In total, 163 clade-specific importation or travel-linked detection events were documented across 41 countries within the primary analytic cutoff; documented secondary transmission occurred in 25 (15.3%) events across 16 countries. Within DRC, 200 health zones recorded 144,035 total reported cases and 2,047 deaths (case fatality ratio 1.42%); these aggregate suspected/clinically compatible and confirmed cases. The top 20 health zones accounted for 45.4% of national burden. Reported transmission declined after the May 2025 peak but persisted beyond PHEIC termination. Emergency termination marked an administrative transition, not the end of mpox transmission. The post-emergency period requires sustained surveillance, maintained diagnostic capacity, targeted response in persistent transmission settings, and equitable access to vaccines and countermeasures.