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

BACKGROUND: Diagnostic 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. OBJECTIVES: To assess the performance (sensitivity/specificity) of molecular assays used by diagnostic laboratories. STUDY DESIGN: Inactivated 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. RESULTS: In 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. CONCLUSIONS: Mpox 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.

Mapping the Mpox discourse: A network and sentiment analysis.

Mpox, a zoonotic disease re-emerging from animals to humans, poses a risk of evolving into a global pandemic due to its high infectivity and potential asymptomatic transmission. This study maps the structure and configuration of mpox-related discussions on Twitter/X, identifies key influencers and top hashtags, and analyzes public sentiment. Data were collected using NodeXL Pro from May 7, 2022, to January 15, 2023, with the keyword "Monkeypox" and visualized using Gephi. Social network analysis ranked nodes by betweenness centrality scores to highlight key communicators, and the YifanHu layout algorithm visualized the network. Influential users, source topics, and hashtags were identified, and sentiment analysis was conducted using Azure Machine Learning tools. The analysis identified 11,397 mpox-related tweets. The network structure resembled a community with diverse participants. Influential users included health and science journalists, writers, academics, medical doctors, and public figures. News media and organizational websites were the top information sources, emphasizing the need for accessible scientific information. "Monkeypox" and "Mpox" were the most prevalent hashtags. Negative sentiments dominated the discussion. This analysis provides insights into network structure, key influencers, information sources, and public sentiment, aiding tailored health initiatives to address misinformation and advocate valid health information and emergency responses.

[Mpox-diagnosis, treatment, immunization, and prognosis]. / Mpox ­ Diagnostik, Therapie, Immunisierung und Prognose.

Mpox (previously known as monkeypox) is receiving attention worldwide due to outbreaks in various countries since May 2022. On August 14, 2024, based on the increase Mpox infections, the World Health Organization (WHO) declared a public health emergency. The zoonotic disease is caused by the Mpox virus, an Orthopoxvirus related to other Poxviridae. The virus is transmitted via direct contact with infected bodily fluids, respiratory droplets, or contaminated objects and has an incubation time of 5-21 days. Symptoms include fever, headache, muscle pain, and a characteristic skin rash which progresses from macules, to papules, to vesicles, and to pustules before scabbing over. There are two main genetic clades of Mpox: clade I (Central Africa) and clade II (West Africa), whereby clade IIb was responsible for the 2022 outbreak. Diagnosis is based on PCR testing of skin lesions. Although Mpox may mimic other diseases such as chickenpox or syphilis, lymphadenopathy is a distinguishing feature. Treatment is primarily supportive, although antiviral agents such as tecovirimat and cidofovir have shown a certain efficacy. Vaccination is an important protective measure; MVA-BN and ACAM2000 are among the available vaccines. Prognosis depends on the clade, the access to medical care, and the underlying health status. Immunocompromised persons and children are at a higher risk of a severe course.

Clinical, epidemiological and molecular aspects of patients with mpox in Romania.

Introduction: To better understand the factors which influence the spread of monkeypox (mpox) infection, the patients that tested positive for mpox virus by real-time PCR in one of the main infectious diseases centers in Bucharest were analyzed in this study, amounting to one third of the confirmed cases in Romania. Methods: Clinical data and laboratory tests were used to build the patient profiles. In the case of positive mpox results, next-generation sequencing of the viral genome was also performed to better comprehend the epidemiology of the infections and the evolutionary path of this virus. Results: Among 47 patients with clinical suspicion of infection, 18 cases tested positive for mpox by real-time PCR (RT-PCR). Patients were mainly men who have sex with men (MSM), often coinfected with HIV-1 (half of the cases) and presenting with other sexually transmitted infections (STIs). Phylogenetic analysis was performed on 20 samples (15 patients) and indicated that mpox cases in Romania were the result of multiple importing events followed by local spread. A few sequences from European countries (Germany, Italy, France) and USA were found to be closely related to the Romanian sequences. Intra-host evolution was observed and documented in one patient with HIV-1 infection with uncontrolled viremia, showing slightly different mutation profiles in two body compartments. Conclusions: This study showed that the mpox cases from Romania presented similar clinical, epidemiological and mutational features with those reported by other European countries.

Mpox Vaccine Acceptance, Democratic Republic of the Congo.

We report general acceptance (61.0%) of an mpox vaccine in the Democratic Republic of the Congo among 5,226 survey respondents. Healthcare workers and respondents in historic mpox-endemic regions had higher acceptance rates. These data highlight the need for increased community engagement and sensitization before widespread deployment of mpox vaccines.

Clade I mpox virus genomic diversity in the Democratic Republic of the Congo, 2018-2024: Predominance of zoonotic transmission.

Recent reports raise concerns on the changing epidemiology of mpox in the Democratic Republic of the Congo (DRC). High-quality genomes were generated for 337 patients from 14/26 provinces to document whether the increase in number of cases is due to zoonotic spillover events or viral evolution, with enrichment of APOBEC3 mutations linked to human adaptation. Our study highlights two patterns of transmission contributing to the source of human cases. All new sequences from the eastern South Kivu province (n = 17; 4.8%) corresponded to the recently described clade Ib, associated with sexual contact and sustained human-to-human transmission. By contrast, all other genomes are clade Ia, which exhibits high genetic diversity with low numbers of APOBEC3 mutations compared with clade Ib, suggesting multiple zoonotic introductions. The presence of multiple clade I variants in urban areas highlights the need for coordinated international response efforts and more studies on the transmission and the reservoir of mpox.

Severe mpox in an immunocompromised patient complicated by deep tissue infection: A case report.

Objectives: We report prolonged mpox (>14 weeks) in a patient with HIV complicated by deep tissue MPXV infection despite two courses of tecovirimat treatment. Methods: MPXV-DNA levels in lesional swabs, blood and tissue were quantified by qPCR. Anti-MPXV antibodies were analyzed by IF and VNT. Infectivity was assessed by virus isolation. Sequencing was performed to assess for tecovirimat resistance mutations and quantitative results were obtained by digital SNP PCR (A288P). Results: The patient's clinical condition improved significantly during both tecovirimat treatment courses (each 14 days), yet we observed persistent MPXV-DNA in lesions accompanied by viremia (mean 1.4 × 104 copies/ml) for >14 weeks. A deep tissue infection driven by MPXV complicated the clinical course (week 9). Presence of infectious virus within the tissue and high infectious titers (>106 PFU/ml) were observed. The VP37 protein sequence revealed A288P substitutions. Digital PCR showed 1 % and less abundance (A288P) during first treatment course (blood and swabs), with increasing proportion during second course (week 8-9; 28 % in blood and swabs), however the mutation was absent in samples from deep tissue infection and MPXV isolates (week 9) indicating compartimentalization. Morphological fully enveloped MPXV partices visualized by TEM in necrotic areas suggesting tecovirimat treatment failure in the deep tissue compartment. Conclusion: Our data provide evidence that Tecovirimat treatment selects for compartimentalized viral mutations (A288P). While the patient clinically benefited from repeated tecovirimat course, emergence of viral muations and deep tissue infection emphasizes the challenge and importance of infectious disease monitoring in mpox patient management.

Comparison of Waning Antibody Responses After Natural Monkeypox Virus Infection and Mpox Vaccination Beyond 6 Months in South Korea.

Natural Monkeypox virus infection induced significantly higher neutralizing antibody titers than Jynneos vaccination, with similar antibody decay rates beyond 6 months. Jynneos recipients with prior smallpox vaccination showed antibody levels comparable to mpox convalescents.

First imported Cases of MPXV Clade Ib in Goma, Democratic Republic of the Congo: Implications for Global Surveillance and Transmission Dynamics.

The ongoing national mpox outbreak in the Democratic Republic of the Congo has resulted in more >30,000 suspected cases in the country from January 2023 to August 2024. While these historic case totals have been driven by primarily by zoonosis, the emergence of Clade Ib monkeypox virus (MPXV), which is connected to more sustained human-to-human transmission, has been associated with increasing public health impacts in eastern DRC. First identified in South Kivu province, Clade Ib MPXV has been identified in multiple non-endemic East African countries for the first time. In DRC, there have been concerns over broader Clade Ib expansion in the country that could further complicate containment and mitigation responses. Here, we report the first introductions of Clade Ib into North Kivu province, including within internal displacement camps, with suspected close contact transmission that includes non-intimate contacts and children. These findings demonstrate that mpox case investigations and community messaging campaigns should include considerations for non-sexual contact-mediated transmission of Clade Ib that includes children <15 years.

MVA-BN vaccine effectiveness: A systematic review of real-world evidence in outbreak settings.

BACKGROUND: Mpox is a disease endemic to Central and West Africa. It caused outbreaks in non-endemic countries, mainly in 2022. The endemic Democratic Republic of Congo is currently experiencing its largest outbreak yet. The vaccine Modified Vaccinia Ankara-Bavarian Nordic (MVA-BN) is approved for active immunization against mpox and smallpox. Since the outbreak in 2022, real-world studies have assessed MVA-BN's vaccine effectiveness (VE) against mpox, and this systematic literature review aims to summarize the most current evidence. METHODS: Medline (via PubMed), Embase, and LILACS were searched, as well as grey literature sources and publications' bibliographies to identify observational studies published between 1/Jan/2022 and 28/Feb/2024 that estimate the VE of MVA-BN against mpox or provide risk measures that allow calculation of these VE estimates. Data were presented descriptively in tables and text; the methodological quality of included records was assessed using NHLBI/NIH quality assessment tools. RESULTS: The literature search identified 16 records that fit the inclusion criteria. The studies took place in high-income countries and were heterogeneous in design, setting, and definition of at-risk populations. MVA-BN VE estimates against mpox infection were assessed. Where the study population was exclusively or primarily those receiving pre-exposure prophylactic vaccination, the adjusted VE estimates ranged from 35 % to 86 % (n = 8 studies) for one dose and from 66 % to 90 % (n = 5) for two doses. Where only post-exposure prophylactic vaccination was assessed, adjusted VE estimates were reported for one dose only at 78 % and 89 % (n = 2). Additionally, MVA-BN reduced the risk of mpox-related hospitalization in one study and the severity of mpox clinical manifestations in two studies. CONCLUSIONS: Despite heterogeneity in study design, setting, and at-risk populations, the reported VE estimates against mpox infection demonstrated the effectiveness of one or two doses of MVA-BN in the context of an outbreak across multiple countries.

Epidemiological analysis of confirmed mpox cases, Burundi, 3 July to 9 September 2024.

We analysed mpox cases in Burundi from July to September 2024, following the introduction of Clade Ib virus. Of 607 samples from the whole population of suspected cases, 154 were PCR-positive, of whom 85 were children under 15 years, with a higher proportion of female children testing positive. Geographical analysis demonstrates case concentration in Bujumbura Mairie (91/154). Age- and sex-specific interventions, as well as community engagement, are important for outbreak containment, as are targeted public health strategies in Burundi.

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.

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

BACKGROUND: The 2022 Monkeypox virus (MPXV) global outbreak boosted development of multiple serological assays to aid understanding of Mpox immunology. OBJECTIVES: The 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. METHODS: Assay 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. RESULTS: The 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. CONCLUSION: Overall, 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.

A qualitative study of Chicago gay men and the Mpox outbreak of 2022 in the context of HIV/AIDS, PrEP, and COVID-19.

During the 2022 mpox outbreak, our study conducted 30 interviews with a recruited sample of Chicago gay men (age 18+) during June-September to investigate their experiences of mpox, HIV/AIDS, and COVID-19. Participants were interviewed with a semi-structured guide about gay sexual identity and social experiences; HIV/AIDS, ART, and PrEP; and COVID-19 behaviors and vaccination. All 30 interview respondents had been fully vaccinated for COVID-19 and expressed minimal COVID-19 vaccine hesitancy. All the men living with HIV in our study were on ART with HIV well controlled. A majority of HIV- participants (70%) were on PrEP, with participants universally aware of PrEP benefits. Additionally, most participants had already received at least one shot of the Jynneos mpox vaccine, with many interviewees enduring long lines, sometimes at multiple locations, before vaccination in primarily gay social spaces. These Chicago gay men demonstrated widespread enthusiasm for mpox vaccination as a disease prevention strategy and most of them had already been vaccinated despite significant barriers. The enthusiasm of the participants in our study emerged within a medical landscape shaped by both COVID-19 vaccination and HIV/AIDS-related health interventions including ART and PrEP, which may have helped instill increased medical trust among this population. Our study suggests that out urban gay men may comprise a distinctive minority population with increased medical trust due to specific social, sexual, and historical experiences.

Characteristics of mpox cases diagnosed in Military Health System beneficiaries, May 2022-April 2024

In May 2022, cases of mpox emerged beyond its historically endemic parts of Central and West Africa, primarily affecting men who have sex with men (MSM), and spreading via sexual networks. During this global outbreak the novel clinical and epidemiological characteristics of mpox disease were thoroughly documented in civilian populations but comparable data have not been reported for Military Health System (MHS) populations including beneficiaries. MHS cases were identified through a variety of data sources, including the Disease Reporting System internet (DRSi) and a customized query of ICD-10-CM (International Classification of Diseases, 10th Revision, Clinical Modification) encounter codes. Contemporaneous chart reviews of patients' electronic health records in the Armed Forces Health Longitudinal Technology Application (AHLTA) and MHS GENESIS were performed to characterize cases. A total of 146 confirmed and probable MHS cases were identified from May 2022 through April 2024. Most cases occurred among MSM, with the majority developing classic prodromal symptoms and some experiencing anogenital and urinary symptoms.

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.