The recent re-emergence of Monkeypox emphasizes the critical importance of ensuring precise and prompt diagnostic capabilities. Traditional diagnosis typically depends on visually examining skin lesions and listening to patients describe their symptoms. However, this approach can be subjective and sometimes leads to misjudgment. To complicate matters further, Monkeypox shares many symptoms with other skin conditions and viral infections, making it difficult for doctors to identify the disease accurately in its early stages. In response, we present Tri-Net, a novel deep learning approach that integrates skin lesion analysis with symptom-based prediction for reliable detection of Monkeypox infection. The skin lesion module leverages an ensemble of three powerful architectures EfficientNetB4, Inception-ResNet V2, and DenseNet201 within the Tri-Net framework to classify images across different stages of the disease. Complementing this, a dedicated Convolutional Neural Network (CNN) processes patient-reported symptoms to further improve diagnostic precision. Our proposed Tri-Net model was trained on publicly available Monkeypox Skin Lesion Dataset (MSLD) comprises diverse dermatological lesion imagery and clinical symptom profiles. The combined approach demonstrated high accuracy, robustness, and significantly outperformed existing methods. Notably, the symptom-based module alone achieved a prediction accuracy of 97.86%. By uniting visual and clinical features, Tri-Net offers a non-invasive, rapid, and highly accurate diagnostic tool that supports early intervention and effective disease control. This study marks a meaningful step toward AI-driven diagnostics for emerging infectious diseases like Monkeypox.

F13 protein is highly conserved in orthopoxviruses and serves as a classic broad-spectrum antiviral target. Nevertheless, clinical trials prove tecovirimat has weak efficacy and poor resistance profile. F13 inhibitors only hinder EEV release to stop viral spread instead of suppressing replication, leaving IMV intact. Current improvements include domain optimization to enhance bioavailability, simplified pharmacophores to boost potency, skeletal editing for better solubility, novel scaffolds for virion assembly inhibition, and dual antiviral-anti-inflammatory agents. Crucially, future paradigms must break away from standalone F13 monotherapy and transition toward combination regimens─such as pairing F13 blockers with host-targeting entry/uncoating inhibitors─or developing multitarget inhibitors that interrupt the viral life cycle prior to IMV maturation by targeting DNA polymerase or transcription machinery.

To expand the supply of the modified vaccinia Ankara (MVA) smallpox/mpox vaccine, fractional doses administered intradermally (ID) have been considered as alternatives to the standard dose given subcutaneously (SC). In a recent clinical trial, participants received the standard 10[8] infectious units of vaccine SC or a one-fifth (IDH) or a one-tenth (IDL) dose ID twice 28-days apart. Two weeks after the second dose, sera were analyzed using a vaccinia virus (VACV) strain Western Reserve plaque-reduction test (PRNT) with the conclusion that IDH but not IDL was non-inferior to SC. The present study extends the latter by quantifying neutralizing antibodies to the MVA vaccine and to monkeypox virus (MPXV) in the absence and presence of added complement and by assessing multiplex binding antibodies to individual VACV antigens (A27, A33, B5, D8 and L1) and the MPXV homologs (A29, A35, B6, E8 and M1). The main conclusions of the study are that the titers of MVA neutralizing antibodies induced by SC and IDH were not significantly different, whereas MVA neutralizing antibody titers induced by IDL were lower like the previous PRNT results. However, the titers of complement-enhanced MPXV neutralizing antibodies induced by SC were significantly higher than for both IDH and IDL immunizations, suggesting that the standard SC dose and reduced dose ID immunizations may not be equivalent. Additional analyses suggested that antibody binding to VACV B5 and D8 and MPXV B6 and E8 may serve as surrogates for neutralizing antibodies.

The A28L protein is involved in mpox virus (MPXV) attachment and fusion, is a major target of neutralizing antibodies, and contains regions prone to genetic variation.

Monkeypox (Mpox) is a viral disease that has garnered global attention due to its human-to-human transmissibility and cross-species transmission. Recent outbreaks in both endemic and non-endemic regions highlight the urgent need to elucidate its molecular mechanisms and evolutionary dynamics. As of now, there are no approved antiviral therapies available for the treatment of Mpox. In this work, we proposed a robust machine learning (ML) and bioinformatics approach to uncover candidate biomarkers associated with Mpox. Two microarray datasets and one RNA-seq dataset were analyzed to screen an initial panel of genes according to p-values. A two-stage feature selection pipeline based on Fisher Score (FS) and Recursive Feature Elimination (RFE) was implemented on the selected gene set. The resulting optimized gene group was subsequently input into our suggested Multilayer Perceptron (MLP) model, which demonstrated superior classification accuracy compared to other classifiers. A total of 33 shared genes were discovered between the differentially expressed genes (DEGs) and the final selected group. Gene Ontology (GO) and KEGG signaling pathway analysis revealed critical biological functions and signaling pathways linked to the common genes. A protein-protein interaction (PPI) network analysis revealed 10 key genes. Regulatory network analysis, incorporating TF-gene and TF-miRNA interactions, highlighted five hub genes with strong associations with regulatory mechanisms. Additionally, molecular docking revealed that ATG3, TRIM14, and DUOX1 exhibited strong interactions with therapeutic compounds. These results indicate that the discovered hub genes may serve as potential candidates for prompt detection, prognosis, and therapeutic targeting in Mpox infection.

We report a prolonged 59-day MPXV infection in a highly immunocompromised HIV-positive patient. We found progressive within-host viral diversification across lesions from different anatomical sites, including the emergence of a mutation previously associated with tecovirimat resistance. These findings highlight the need to investigate alternative therapeutic strategies for MPXV in severely immunocompromised patients.

Human Monkeypox is a smallpox-like zoonotic infection transmitted through animals, contaminated objects, or human contact, posing growing public health risks. Despite its declaration as a global health emergency in 2022, community awareness remains limited in Ethiopia's Oromia region, where ecological and socioeconomic factors heighten vulnerability. Thus, this community-based cross-sectional study, conducted from July 6 to August 30, 2025, among 842 adults in Oromia, assessed knowledge, attitudes, and associated factors regarding Human Monkeypox. Data were analyzed using descriptive statistics and logistic regression in SPSS version 27, with significance set at p < 0.05. Results showed that 56.5% of participants had good knowledge of Human Monkeypox, significantly associated with age 30-40 years, marital status, occupation, income, and access to information through media, health workers, or peers. Furthermore, 57.5% demonstrated a positive attitude, which was strongly linked to higher knowledge levels, while lower education and farming occupation reduced positivity. Knowledge, shaped by education, occupation, age, and information access, was a strong determinant of positive attitudes. Overall, limited awareness and negative perceptions highlight the need for community-based, culturally sensitive educational strategies leveraging accessible information channels to enhance preparedness and promote preventive practices against Human Monkeypox.

Monkeypox (Mpox) is a zoonotic disease caused by MPXV and remains a threat to global public health. MPXV Clade Ib has higher mortality, whereas Clade IIb exhibits widespread international transmission. WHO has twice designated mpox epidemics as a Public Health Emergency of International Concern (PHEIC). This highlights that the virus is not only pathogenic but also has the potential to cause a global pandemic, posing a challenge to the international public health system. The rapid transmission of MPXV between and within hosts is related to two key virus particles: IMV and EEV. Antigenic proteins (A29L, M1R, B6R, and A35R) on these particles mediate viral invasion, dissemination, and immune evasion. Clinical management of mpox remains supportive, and there is a lack of specific antiviral drugs. Although smallpox-related drugs (such as tecovirimat, cidofovir, and brincidofovir) show potential, their clinical efficacy needs to be verified. The existing vaccines recommended by the WHO are facing challenges, such as side effects or limited duration of immune protection in application, prompting the research and development of novel vaccines and antibody therapeutics. Research on the mechanism of invasion and surface proteins of viruses will continue to provide a scientific foundation for the development of medications and vaccines. Prevention and control rely on a stronger monitoring system, timely risk warnings, and public health education. This article reviews the recent progress in MPXV detection, vaccine development, and antibody-based therapy, providing new insights for mpox prevention and control.

Conventional polymerase chain reaction (PCR)-based detection methods suffer from time-consuming procedures, reliance on specialized equipment, and difficulty in achieving early viral diagnosis. In this study, interferometric fiber-optic sensing is integrated with the CRISPR/Cas12a system for the first time. With sensitivity further enhanced by immobilizing ZnO@Au on the fiber surface, the platform enables rapid, amplification-free detection of monkeypox virus (MPXV) at the single-molecule level. Whispering-gallery modes (WGMs) excited in the fiber probe provide high sensitivity to ambient refractive-index changes, while the ZnO@Au layer induces localized surface plasmon resonance (LSPR) and coupled plasmon-waveguide resonance (CPWR) on the fiber surface. By controlling the AuNPs occupancy on ZnO, the LSPR and CPWR absorption peaks can be tuned to match the demodulation spectral band. Moreover, the ZnO-Au heterojunction further strengthens the LSPR, thereby improving the sensitivity of the fiber probe. The resulting sensing probe achieves amplification-free detection of plasmid targets from both MPXV subtypes down to 10° copies/μL, with the entire assay completed within 9 min. The detection capability was validated using real clinical MPXV samples, showing complete agreement with qPCR results. The strategy proposed in this work offers a feasible approach for early and rapid viral detection.

The monkeypox virus (MPXV) belongs to the Poxviridae family and has emerged as a global problem for public health since May 2022. Both innate and adaptive immunity are crucial for preventing MPXV infections. Understanding the interaction between the innate and adaptive immune systems and MPXV could be critical in developing novel treatments. This review presents an update on the host antiviral innate and adaptive immune responses against four zoonosis poxvirus, including mouse kidney parvovirus, cowpox virus, variola virus, and vaccinia virus. The effect of APOBEC3 variants on viral immune escape and the underlying mechanisms were also explored. The possible mechanisms of the effects of human immunodeficiency virus and monkeypox co-infection in the clinical setting were also analysed in the hope of providing a basis for clinical drug development and treatment.

The ongoing spread of the monkeypox outbreak underscores the urgent need for the development of effective therapeutics against the monkeypox virus (MPXV). Aminopterin is a classical antifolate agent known for its anti-tumor activity through the inhibition of folate metabolism. Here, aminopterin was identified as an effective anti-MPXV agent. It significantly inhibited MPXV replication, unlike other antifolate agents such as methotrexate, pemetrexed, and LSN3213218. In Vero E6 cells, the half-maximal inhibitory concentration (IC50) was 36.10 nM, the half-cytotoxic concentration (CC50) was 3,310 nM, and the selectivity index (SI) was 91.690. In human foreskin fibroblast cells, the IC50 was 32.73 nM, CC50 exceeded 10 μM, and SI was greater than 305.530. In vivo, aminopterin not only inhibited MPXV infection but also mitigated high-dose viral infection-associated inflammation. Furthermore, the data indicated that aminopterin reduced viral load by interfering with viral attachment and replication. Together, these findings identify aminopterin as a promising candidate for anti-MPXV drug development.