Monkeypox Virus Research Articles

Wheat Germ Agglutinin-Modified "Three-in-One" Multifunctional Probe Driven Broad-Spectrum and Flexible Immunochromatographic Diagnosis of viruses With High Sensitivity.

The conventional lateral flow assay (LFA) fails to the demands for the accurate screening of viruses as a result of its low sensitivity of colorimetric signal output and poor universality limited by antibody pairs. Here, a magnetically assisted dual-signal output LFA platform is developed for the ultrasensitive, universal, and flexible detection of viruses. A "three-in-one" multifunctional probe (MAuDQD) is prepared using a 180nm Fe3O4 core to load numerous Au nanoparticles (NPs) and two layers of QDs, which can substantially improve the sensitivity of LFA through coupling with the effects of magnetic enrichment and colorimetric/fluorescent enhancement. Wheat germ agglutinin (WGA)-modified MAuDQD attained the broad-spectrum capture viral membrane proteins and the colorimetric/fluorescent dual-mode detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and monkeypox virus (MPXV) on the LFA strip. In the colorimetric mode, the target viruses detected directly, with the visual sensitivity reaching 0.1-0.5ngmL-1 and the fluorescent mode supported quantitative analysis of SARS-CoV-2/MPXV with limits of detection decreasing to pgmL-1 level. Practicability of the MAuDQD@WGA-LFA is verified through the detection of 33 real clinical samples, showing the proposed assay has a great potential to become a sensitive, accurate, and universal tool for on-site monitoring of viral infections.

International external quality assessment study for detection of monkeypox virus by PCR supporting laboratory preparedness during the 2022–2023 mpox outbreak and beyond

BackgroundDiagnostic capabilities and correspondent External Quality Assessments (EQA) are key for outbreak preparedness. To support diagnostic facilities with a quality assessment of newly established monkeypox virus (MPXV) molecular diagnostic workflows, Quality Control for Molecular Diagnostics (QCMD) and the Bundeswehr Institute of Microbiology (IMB) piloted an international EQA study conducting four challenges from autumn 2022 to summer 2023 during the global mpox outbreak. ObjectivesTo assess the performance (sensitivity/specificity) of molecular assays used by diagnostic laboratories. Study designInactivated EQA panels were prepared and distributed containing seven samples of clade Ia and clade IIb MPXV strains at different viral concentrations, two specificity controls with other zoonotic orthopoxviruses (vaccinia and cowpox virus) and a negative control. Assessment was based on reported qualitative testing results. ResultsIn this outbreak-related EQA study, a total of 192 laboratories from 37 countries reported 346 qualitative datasets. Overall, core samples were correctly detected by approximately 92 % of participants in all four challenges. While sensitivity performance was acceptable in at least 90 % of datasets correctly reported even for educational MPXV-positive samples with low viral concentration [102 genome equivalents (GE)/mL], several laboratories reported the educational specificity controls as false positives or were unable to differentiate MPXV from related orthopoxviruses. ConclusionsMpox is now a globally occurring infection with a demand for quality-assured diagnostic capabilities. The newly established EQA scheme presented here, offers a multi-purpose panel for orthopoxviruses with a focus on MPXV which will continue to ensure diagnostic quality in clinical settings with up-to-date sample panels.

Evaluation of a multiplexed immunoassay for assessing long-term humoral immunity Orthopoxviruses

BackgroundThe 2022 Monkeypox virus (MPXV) global outbreak boosted development of multiple serological assays to aid understanding of Mpox immunology. ObjectivesThe study aimed to assess a multiplexed solid-phase electrochemiluminescence immunoassay (Meso Scale Discovery (MSD)) for simultaneous detection of antibodies against MPXV, including A35, E8 and M1 antigens, along with corresponding Vaccina Virus (VACV) homologues and demonstrate its accuracy in assessing antibody titres post-vaccination and infection. MethodsAssay performance was assessed for simultaneous detection of antibodies against MPXV and corresponding VACV antigens. Sensitivity and specificity were evaluated with paediatric negatives (n = 215), pre- and post-IMVANEX vaccinated (n = 80), and MPXV (Clade IIb, n = 39) infected serum samples. ResultsThe assay demonstrated high specificity (75.68 % (CI: 69.01–81.29) - 95.98 % (CI:92.54–97.87)) and sensitivity (62.11 % (CI:52.06–71.21) - 98.59 % (CI:92.44 %–99.93 %)) depending on the Orthopoxvirus antigen. Preferential binding was observed between MPXV-infected individuals and MPXV antigens, while vaccinated individuals exhibited increased binding to VACV antigens. These results highlight differential binding patterns between antigen homologues in related viruses. ConclusionOverall, this assay demonstrates high sensitivities in detecting antibodies for multiple relevant MPXV and VACV antigens post-infection and post-vaccination, indicating its utility in understanding immune responses to Orthopoxviruses in current and future outbreaks and evaluating the immunogenicity of new-generation Mpox-specific vaccinations.

MPXV Infection Stimulates a More Robust and Durable Neutralizing Antibody Response Compared to MVA-BN Vaccination.

Monkeypox virus (MPXV) has recently caused a global disease outbreak in humans. Differences in the neutralizing antibody response to vaccination vs. MPXV infection remain poorly understood. Here, we examined the neutralization of MPXV and VACV by sera from a cohort of convalescent and vaccinated individuals at 1- and 8-months post-exposure. Convalescent individuals displayed higher neutralizing antibody titers against MPXV than vaccinated and MPXV-naïve persons at one-month post-exposure. Neutralizing antibody titers had waned significantly in both groups at 8 months. This study suggests additional vaccine strategies are needed to elicit a durable humoral response and prevent breakthrough infections.

Impact of surveillance in human-to-human transmission of monkeypox virus

Impact of surveillance in human-to-human transmission of monkeypox virus

Monkeypox Clade Ib virus introduction into Burundi: first findings, July to mid-August 2024.

We describe cases with monkeypox virus (MPXV) Clade Ib in Burundi from their first detection in July until 20 August 2024. Testing 442 people with vesicular lesions confirmed 170 cases (98 male; 72 female), 82 (48%) being < 15 years old. Differential diagnosis of the first 30 individuals testing MPXV negative revealed chickenpox in 20. Cases occurred in 26 of 49 Burundi health districts, but mostly in Bujumbura Nord (88/170; 67%). Case-derived MPXV genetic sequences from Burundi and South-Kivu (Democratic Republic of the Congo), clustered together in phylogenetic analysis.

Diagnostic and surveillance testing capability for mpox in the EU/EEA, September 2024.

In response to the increasing number of mpox cases caused by monkeypox virus (MPXV) clade I in the African continent and the first reported travel-related clade Ib case of mpox in EU/EEA, the European Centre for Disease Prevention and Control surveyed national capability for detection and characterisation of MPXV in the EU/EEA. The results showed high level of capability for case confirmation by PCR, alongside molecular typing methods for identification of MPXV clades and/or clade I subclades within the EU/EEA.

Mpox and Lessons Learned in the Light of the Recent Outbreak: A Narrative Review.

According to the WHO, more than 90,000 cases of mpox have been reported since the 2022 worldwide outbreak, which resulted in 167 deaths, while a new outbreak in Africa since 2023 has resulted in over 18,000 cases and 617 deaths. Mpox is a zoonosis caused by the monkeypox virus, a double-stranded DNA virus belonging to the Orthopoxvirus genus, which causes smallpox-like illness. Until 2022, cases were predominately located in West and Central Africa, with only sporadic cases and outbreaks reported in other parts of the world. During the 2022 outbreak, the primary mode of transmission was sexual contact among men who have sex with men. The changing epidemiology of mpox resulted in new disease phenotypes and populations at risk, disproportionally affecting people who live with HIV. Commonly presenting as a mild, self-limiting illness, mpox can cause severe and protracted disease in people with HIV with a CD4 count < 200 cell/mm3. The global emergence of mpox that followed and intersected with COVID-19 mobilized the scientific community and healthcare stakeholders to provide accurate diagnostics, preventive vaccines and treatment to those most affected. Despite existing gaps, this rapid response helped to contain the outbreak, but challenges remain as new variants emerge. Preparedness and readiness to respond to the next outbreak is crucial in order to minimize the impact to the most vulnerable.

Knowledge Regarding Human Monkey pox (HMPX) Among Nursing Students

Monkey pox is a viral infectious disease caused by the monkey pox virus, which belongs to the same family as the virus causing smallpox. Though typically self-limiting, with symptoms lasting two to four weeks, it can be transmitted through close contact with infected individuals, animals, or contaminated materials. Materials and Methods: A pre-experimental one-group pretest post-test designed study was conducted to evaluate the effectiveness of Laptop-Assisted Teaching on knowledge regarding Human Monkey pox (HMPX) among 60 nursing students using purposive sampling technique at a selected school in Bengaluru with the Objectives, to assess the effectiveness of Laptop-Assisted Teaching in improving knowledge about Human Monkey pox (HMPX) among nursing students and to determine the association between pretest knowledge scores regarding Human monkey pox (HMPX) among Nursing students with their selected sample characteristics. Data was collected using a structured knowledge questionnaire on HMPX, validated by 10 experts. Descriptive and inferential statistics were used for analysis. Results: The study showed that Laptop-Assisted Teaching significantly improved students' knowledge of HMPX. The mean pretest score was 9.95 (SD = 4.01), while the mean post-test score was 17.84 (SD = 8.78). The computed paired ‘t’ test value of 6.36 at p&gt;0.05 was significant. Additionally, there was a statistically significant association between pretest knowledge scores and selected sample characteristics such as age and family type. Conclusion: The study concluded that nursing students had inadequate knowledge about HMPX initially, but Laptop-Assisted Teaching was effective in enhancing their understanding.

MSMP-Net: A Multi-Scale Neural Network for End-to-End Monkeypox Virus Skin Lesion Classification

Monkeypox is a zoonotic disease caused by monkeypox virus infection. It is easily transmitted among people and poses a major threat to human health, making it of great significance in public health. Therefore, this paper proposes MSMP-Net, a multi-scale neural network for end-to-end monkeypox virus skin lesion classification ConvNeXt is used as the backbone network, and designs such as inverse bottleneck layers and large convolution kernels are used to enhance the network’s feature extraction capabilities. In order to effectively utilize the multi-level feature maps generated by the backbone network, a multi-scale feature fusion structure was designed. By fusing the deepest feature maps of multi-scale features, the model’s ability to represent monkeypox image features is enhanced. Experimental results show that the accuracy, precision, recall, and F1-score of this method on the MSLD v2.0 dataset are 87.03 ± 3.43%, 87.59 ± 3.37%, 87.03 ± 3.43%, and 86.58 ± 3.66%, respectively.

High-Throughput Insilico Drug Screen against Mpox Targeted Proteins in Comparison with Repurposed Antiviral Drugs against Natural Compounds

The recent resurgence of the Monkeypox Virus (MPXV) has prompted increased efforts to discover effective antiviral treatments. This study utilized an in silico docking approach with CB Dock2 to assess both natural compounds and repurposed antiviral medications. We concentrated on several critical viral targets for inhibition, specifically VP39 2'-O Methyltransferase, viral topoisomerase-DNA complexes, and poxin. Our results identified a number of natural substances, including Physalin A, Sitoindoside IX, Withanolide, Shatavarin 1, Kutkoside, and Berberine HCl, as promising inhibitors. Tecovirimat was found to be the most effective among the repurposed antivirals. Notably, natural products like Withanolide, Sitoindoside IX, and Physalin A showed significant inhibitory potential based on their Vina scores and binding affinities, suggesting they may serve as alternative therapeutic options. Tecovirimat consistently proved to be the most powerful inhibitor among repurposed antivirals across all examined targets, reinforcing its importance in combating MPXV.

Knowledge and conspiracy belief of healthcare professionals on human monkeypox virus: a cross-sectional study in a tertiary hospital of Bangladesh

Background: Since early May 2022, the unprecedented outbreak of human monkey pox (HMPX) has been documented globally which demands assessment of knowledge on the emerging viral infection as well as the state of conspiracy belief among the healthcare professionals. The present study aimed to assess the knowledge about HMPX and its relation to conspiracy belief. Methods: This cross-sectional study was conducted at the Combined Military Hospital Dhaka (CMH) among purposively selected 254 healthcare professionals from July 2022 to September 2022. Data were collected through face-to-face interview using a pre-tested semi-structured questionnaire with validated and reliable tools. Results: The study comprised a total of 254 respondents, including 53.9% male, 72.8% respondents having a graduate degree and 42.9% individuals aged between 26-30 years. The study revealed very poor (61.8%) knowledge on HMPX as well as higher (50.4%) belief in conspiracy theory among the respondents. The hierarchical logistic regression revealed that male had significantly higher (B=1.12, OR 3.06, 95% CI=1.714-5.463) knowledge on HMPX virus than that of the female, age group &lt;31 years emerged as a negative and significant predictor than that of the age group &gt;31 years (B= -1.065, OR 0.345, 95% CI=0.172-0.690), monthly income of between 60001-90000 taka also emerged as a positive and significant predictor than that of the income of &gt;90001 taka (B=0.763, OR 2.145, 95% CI=1.002-4.595). Conclusion: The study findings demand intervention of appropriate health educational program in the medical study curriculum. JOPSOM 2023; 42(1):29-39

Notes from the Field: Mpox Cluster Caused by Tecovirimat-Resistant Monkeypox Virus - Five States, October 2023-February 2024.

The antiviral drug tecovirimat* has been used extensively to treat U.S. mpox cases since the start of a global outbreak in 2022. Mutations in the mpox viral protein target (F13 or VP37) that occur during treatment can result in resistance to tecovirimat† (1,2). CDC and public health partners have conducted genetic surveillance of monkeypox virus (MPXV) for F13 mutations through sequencing and monitoring of public databases. MPXV F13 mutations associated with resistance have been reported since 2022, typically among severely immunocompromised mpox patients who required prolonged courses of tecovirimat (3-5). A majority of patients with infections caused by MPXV with resistant mutations had a history of tecovirimat treatment; however, spread of tecovirimat-resistant MPXV was reported in California during late 2022 to early 2023 among persons with no previous tecovirimat treatment (3). This report describes a second, unrelated cluster of tecovirimat-resistant MPXV among 18 persons with no previous history of tecovirimat treatment in multiple states.

Therapeutic agents for the treatment of human mpox.

The aim of this study was to summarize the current knowledge of therapeutic options for mpox (formerly known as monkeypox) in the context of recent outbreaks and the ongoing evolution of the virus. Multiple therapeutic agents, including tecovirimat, cidofovir, brincidofovir, and vaccinia immune globulin, have been used during the multicountry outbreak of mpox caused by Clade 2b monkeypox virus that began in 2022. Tecovirimat has been most extensively used, based on efficacy against mpox lethal challenge in animal models, and human safety data. Real-world observational evidence has further supported safety with minimal adverse events in large cohorts and mixed reports of reductions in time to lesion resolution. Several prospective randomized controlled trials using tecovirimat are underway with headline results from a study in the Democratic Republic of the Congo showing no difference in lesion resolution compared to placebo. Other studies including in outpatient settings are underway in Europe and the Americas. Cidofovir and brincidofovir, limited by adverse event profiles, have been less extensively studied. Vaccinia immune globulin has been used predominantly in salvage therapy for severe mpox, with no large observational series available. The 2022 multicountry outbreak of mpox marked a public health emergency. Agents approved for smallpox management were widely used for mpox, supported by animal and in-vitro evidence, and human safety data. The large number of human cases has allowed retrospective observational study of these agents and facilitated recruitment in prospective trials. The ongoing evolution of the virus may pose challenges for therapeutic interventions, necessitating rigorous randomized controlled trials to guide clinical use.

Network-Based Drug Repurposing and Genomic Analysis to Unveil Potential Therapeutics for Monkeypox Virus.

The emergence of the human monkeypox virus (MPXV) and the lack of effective medications have necessitated the exploration of various strategies to combat its infection. This study employs a network-based approach to drug discovery, utilizing the BLASTn and phylogenetic analysis to compare the MPXV genome with those of 18 related orthopoxviruses, revealing over 75 % genomic similarity. Through a literature review, 160 human-host proteins linked to MPXV and its relatives were identified, leading to the construction of a human-host protein interactome. Analysis of this interactome highlighted 39 central hub proteins, which were then examined for potential drug targets. The process successfully revealed 15 targets already approved for use with medications. Additionally, the functional enrichment analysis provided insights into potential pathways and disorders connected with these targets. Four medications, namely Baricitinib, Infliximab, Adalimumab, and Etanercept, have been identified as potential candidates for repurposing to combat MPXV. In addition, the pharmacophore-based screening identified a molecule that is comparable to Baricitinib and has the potential to be effective against MPXV. The findings of the study suggest that ZINC22060520 is a promising medication for treating MPXV infection and proposes these medications as potential options for additional experimental and clinical assessment in the battle against MPXV.

Unleashing thymidylate kinase inhibitors: a computational quest against monkeypox virus

Unleashing thymidylate kinase inhibitors: a computational quest against monkeypox virus

Clinicopathological study of 24 cases of monkeypox virus infection-related rashes

Objective: To investigate the clinicopathological characteristics of rashes in monkeypox patients through a series of skin biopsies, and examine their pathological features and the most effective tests. Methods: Patients with monkeypox virus infection admitted to Beijing Ditan Hospital from June to August 2023 were identified. Among them, 24 patients underwent skin biopsies for clinical pathological study that were included in this study. Clinical information, rash pictures, and nucleic acid test results were analyzed using histopathology, immunohistochemistry, RNAscope® hybridization and electron microscopy. Results: All 24 patients were male, including 14 patients with concurrent human immunodeficiency virus infection. Their average age was (32.3±5.4) years. The nucleic acid test confirmed monkeypox virus infection. The clinical feature of monkeypox rashes was solitary rather than clustered distribution, with rashes occurring in similar phase, distinguishing it from herpesvirus. The rashes in these patients were mostly scattered, with an average of (13.0±11.8) rashes, and most commonly present in the perineum, face, limbs, and trunk. The three main pathological features of these rashes were ballooning degeneration of the epidermal spinous cell layer, the characteristic intra-cytoplasmic Guarnieri's bodies and significant infiltration of inflammatory cells in whole dermal layer. Immunohistochemistry, RNAscope® hybridization, and electron microscopy can all effectively detect the monkeypox virus. Electron microscopy showed viral replication in various types of skin cells. Conclusions: The study describes the pathological features of monkeypox virus rashes. Pathological examination of skin biopsy samples is helpful to diagnose these rashes. The study suggests that the monkeypox virus has a unique epitheliotropic affinity and can infect various types of cells in the skin.

A review of Mpox: Biological characteristics, epidemiology, clinical features, diagnosis, treatment, and prevention strategies

AbstractThe outbreak of monkeypox virus (MPXV) was declared a Public Health Emergency of International Concern (PHEIC) by the World Health Organization (WHO), and the zoonotic disease caused by viral infection was renamed as “Mpox” on November 28, 2022. Currently, there is no approved vaccine or specific antiviral treatment for Mpox, and a main preventive strategy against MPXV infection remains the smallpox vaccine. Although there was an emergency use authorization (EUA) of Brincidofovir and Tecovirimat for the clinical treatment of clade II Mpox, while Tecovirimat failed to reduce the duration of Mpox lesions among patients infected with clade I Mpox in the Democratic Republic of the Congo (DRC). Therefore, it is still an urgent need to develop an effective medication. This review aims to enhance the understanding of Mpox and contribute to its prevention and treatment strategies, it provides a systemic introduction of the biological and epidemiological characteristics of MPXV, the clinical feature and diagnosis of Mpox, as well as treatment and prevention strategies, which will improve the comprehension about MPXV and offer potential strategies for clinical treatment.

Bioinformatic identification of monkeypox virus phylogenetic gene trees that are representative of its whole-genome phylogenetic tree.

Phylogenetic analysis based on whole-genome sequences is the gold standard for monkeypox virus (MPXV) phylogeny. However, genomic epidemiology capability and capacity are lacking or limited in resource poor countries of sub-Saharan Africa. Therefore, these make real-time genome surveillance of MPXV virtually impossible. We hypothesized that phylogenetic analysis based on single, conserved genes will produce phylogenetic tree topology consistent with MPXV whole-genome phylogeny, thus serving as a reliable proxy to phylogenomic analysis. In this study, we analyzed 62 conserved MPXV genes and showed that Bayesian phylogenetic analysis based on five genes (OPG 066/E4L, OPG068/E6R, OPG079/I3L, OPG145/A18R, and OPG150/A23R) generated phylogenetic trees with 72.2-96.3% topology similarity index to the reference phylogenomic tree topology. Our results showed that phylogenetic analysis of the identified five genes singly or in combination can serve as surrogate for whole-genome phylogenetic analysis, and thus obviates the need for whole-genome sequencing and phylogenomic analysis in regions where genomic epidemiology competence and capacity are lacking or unavailable. This study is relevant to evolution and genome surveillance of MPXV in resource limited countries.

Viral genetics and transmission dynamics in the second wave of mpox outbreak in Portugal and forecasting public health scenarios

ABSTRACT In 2023, a second wave of the global mpox epidemic, which is mainly affecting men who have sex with men (MSM), was observed in some countries. Herein, we benefited from a large viral sequence sampling (76/121; 63%) and vast epidemiological data to characterise the re-emergence and circulation of the Monkeypox virus (MPXV) in Portugal during 2023. We also modelled transmission and forecasted public health scenarios through a compartmental susceptible-exposed-infectious-recovered (SEIR) model. Our results suggest that the 2023 mpox wave in Portugal resulted from limited introduction(s) of MPXV belonging to C.1.1 sublineage, hypothetically from Asia, followed by sustained viral transmission and potential exportation to other countries. We estimated that the contribution of the MSM high sexual activity group to mpox transmission was 120 (95% CrI: 30–3553) times higher than that of the low sexual activity group. However, among the high sexual activity group, vaccinated individuals likely contributed approximately eight times less [0.123 (95% CrI: 0.068–0.208)] than the unvaccinated ones. Vaccination was also linked to potential reduced disease severity, with a Mpox Severity Score of 6.0 in the vaccinated group compared to 7.0 in unvaccinated individuals. Scenario analysis indicated that transmission is highly sensitive to sexual behaviour, projecting that a slight increase in the MSM sub-population with high sexual activity can trigger new mpox waves. This study strongly supports that continued vaccination, targeted awareness among risk groups and routine genomic epidemiology is needed to anticipate and respond to novel MPXV threats (e.g. global dissemination of clade I viruses).