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- Demographic and Clinical Characteristics of Mpox Patients Attending an STD Clinic in LisbonPublication . Cid Brito, Margarida; Nuncio, M.S.; Lopes de Carvalho, Isabel; Cordeiro, Rita; Pelerito, AnaMpox is a viral disease caused by the monkeypox virus, which marked the year of 2022 with a global outbreak. While previously considered to be a zoonosis of almost exclusive animal-to-human transmission, the current outbreak has been attributed to human-to-human transmission, particularly sexual transmission. As a new sexually transmissible disease, we studied the epidemiological and clinical features, as well as the concomitant occurrence of other sexually transmissible diseases, treatment approach, and outcome of our 291 patients, in the current outbreak. We found a total of 169 concomitant sexually transmissible infections of bacterial and viral origins, corresponding to 107 patients. Neisseria gonorrhoeae was the most common agent, particularly in the anal location. With this work, we emphasize the need for a thorough epidemiological and medical history, as well as a concomitant complete laboratorial screening for other STIs in patients with confirmed or suspected mpox.
- Coordinated implementation of a conventional PCR assay to detect all Ebola and Marburg virus species in a European laboratory networkPublication . Heimsch, K.C.; Bleicker, T.; Best, T.D.; Presser, L.D.; Molenkamp, R.; Jääskeläinen, A.J.; Milewska, A.; Smahelová, J.; Baronti, C.; Pappa, S.; Tabain, I.; Cordeiro, Rita; Marsili, G.; Huik, K.; dos Reis, V. Pinho; Barzon, L.; Maes, P.; Drosten, C.; Corman, V.M.Background: Filoviruses, including Ebola and Marburg viruses, cause severe hemorrhagic fever in humans and primates. These viruses pose significant threats to public health, making rapid and sensitive detection critical for controlling outbreaks. We developed and validated a hemi-nested generic PanFilo assay to detect all Ebola virus species, Marburg viruses, and recently discovered bat filoviruses. This assay was deployed to 15 European laboratories and evaluated through testing of eight non-infectious samples. Objectives: Laboratories were asked to determine the detection limit of positive controls and test all samples using the assay provided. The deployed assay enables direct Nanopore sequencing of PCR products, by using tagged primers during the second round of PCR. Sequencing of the samples was carried out on a voluntary basis. Results: Multicenter validation revealed a 95 % limit of detection of 5309 RNA copies/μL for Ebola, 10,273 copies/μL for Marburg, and 2145 copies/μL for Mengla virus. In an implementation quality assessment, 93.3 % (84/90) of samples containing filovirus RNA were correctly identified and 100 % (30/30) of filovirus-negative samples were correctly identified. Thirteen laboratories sequenced PCR products, with nine identifying all positive samples correctly. Conclusion: The assay enables rapid and reliable detection of filoviruses, with sequencing capabilities for identifying both known and novel variants. This assay might be used for detection during the initial phase of an emerging filovirus outbreak, before a specific assay has been developed. However, our distribution across 15.
- The impact of orthopoxvirus vaccination and Mpox infection on cross-protective immunity: a multicohort observational studyPublication . Crandell, Jameson; Pischel, Lauren; Fang, Zhenhao; Conde, Luciana; Zhong, Yi; Lawres, Lauren; Meira de Asis, Gustavo; Maciel, Gabriela; Zaleski, Agnieszka; Lira, Guilherme S.; Higa, Luiza M.; Breban, Mallery I.; Vogels, Chantal B.F.; Caria, João; Pinto, Ana Raquel; Almeida, Vasco; Maltez, Fernando; Cordeiro, Rita; Póvoas, Diana; Grubaugh, Nathan D.; Aoun-Barakat, Lydia; Grifoni, Alba; Sette, Alessandro; Castineiras, Terezinha M.; Chen, Sidi; Yildirim, Inci; Vale, Andre M.; Omer, Saad B.Background: Cross-reactive immune memory responses to orthopoxviruses in humans remain poorly characterised despite their relevance for vaccine design and outbreak control. We aimed to assess the magnitude, specificity, and durability of cross-reactive immune responses elicited by smallpox vaccines and mpox virus infection. Methods: We did a multicohort observational study involving participants from the USA, Brazil, and Portugal across four groups: Dryvax (first-generation smallpox vaccine) recipients vaccinated 40-80 years ago, JYNNEOS (third-generation smallpox vaccine) recipients vaccinated within the past year, a cohort receiving both vaccines, and patients infected with clade IIb mpox. Samples were analysed for systemic and mucosal humoral responses, neutralising antibody titres, viral antigen structural analysis, and T-cell cross-reactivity to vaccina virus, cowpox virus, and mpox virus. Statistical analyses included correlation assessments and comparisons across cohorts to determine the magnitude, longevity, and breadth of immune responses. Findings: Between July 7, 2022, and Aug 3, 2023, 262 participants were recruited, resulting in analysis of 378 samples. Both first-generation and third-generation smallpox vaccines elicited vaccinia virus-reactive and mpox virus-reactive antibodies, with the strongest responses targeting the less conserved extracellular virion antigens B5 and A33. Despite high concentrations of anti-mpox virus antibodies in the plasma, cross-neutralisation activity correlated with viral antigenic distance. Higher neutralisation was observed for cowpox virus than for mpox virus, which has lower antigenic conservation with vaccina virus. Complement-mediated neutralisation enhanced mpox virus neutralisation, overcoming the limitations of antigenic distance. Dryvax recipients sustained vaccina virus neutralisation titres for over 80 years, whereas cross-reactive responses did not show this durability. JYNNEOS-induced responses waned within a year. T-cell cross-reactivity was long-lasting, detected up to 70 years after vaccination. Booster vaccinations augmented the magnitude, breadth, and longevity of cross-neutralising responses. Interpretation: Our findings highlight the potential combined role of antibody effector functions and T-cell memory in cross-protection against orthopoxviruses. Complement-mediated neutralisation enhances cross-protection, overcoming antigenic distance. These Fc-mediated functions, along with T-cell responses, contribute to effective and long-lasting immunity conferred by smallpox vaccines against other orthopoxviruses.
- Mpox in People Living with HIV: Clinical Challenges, Preventive Strategies and Public Health ImplicationsPublication . Cordeiro, Rita; Caria, J.; Sobral, D.; Póvoas, D.; .Monkeypox virus (MPXV) re-emerged in 2022 with a global outbreak that affected more than 100,000 individuals worldwide. People living with HIV (PLWH) accounted for a substantial proportion of cases, raising concerns about disease presentation, management, and outcomes in this population. Evidence indicates that PLWH with advanced or uncontrolled HIV infection experienced more severe mpox, with higher hospitalization rates, more complications, and longer disease courses. In contrast, individuals with well-controlled HIV generally had outcomes similar to those without HIV. Access to timely diagnosis, consistent antiretroviral therapy, and availability of tecovirimat were key factors influencing prognosis. Reports also suggest bidirectional interactions between mpox and HIV pathogenesis. Immune activation and APOBEC3-related viral evolution have been proposed; however, these mechanisms remain incompletely characterized and warrant further investigation. Moreover, disparities in healthcare access and stigma compound the vulnerability of PLWH, emphasizing the need for integrated approaches.
- Mpox Surveillance and Laboratory Response in Portugal: Lessons Learned from Three Outbreak Waves (2022-2025)Publication . Cordeiro, Rita; Francisco, Rafaela; Pelerito, Ana; Lopes de Carvalho, Isabel; Nuncio, MSBackground/Objectives: Mpox re-emerged in 2022 as a global health concern. Between 2022 and 2025, Portugal experienced three distinct outbreak waves, highlighting the critical role of laboratory surveillance and public health interventions. This study describes the epidemiological trends, diagnostic performance, and key lessons learned to improve outbreak preparedness. Methods: A total of 5610 clinical samples from 2802 suspected cases were analyzed at the National Institute of Health Doutor Ricardo Jorge using real-time PCR methods. Positivity rates and viral loads (Ct values) were assessed across different clinical specimen types, including lesion, anal, oropharyngeal swabs, and urine samples. Results: Mpox was confirmed in 1202 patients. The first outbreak accounted for 79.3% of cases (n = 953), followed by a significant reduction in transmission during subsequent waves. Lesion and rectal swabs provided the highest diagnostic sensitivity (95.1% and 87.9%, respectively). Oropharyngeal swabs contributed to diagnosis in cases without visible lesions, while urine samples showed limited utility. Conclusions: This study underscores the importance of sustained laboratory surveillance and adaptive public health strategies in controlling mpox outbreaks. Optimizing specimen collection enhances diagnostic accuracy, supporting early detection. Continuous monitoring, combined with targeted vaccination and effective risk communication, is essential to prevent resurgence and ensure rapid response in non-endemic regions.
- Undetected circulation of monkeypox virus in Portugal: Evidence for a 50-day gap before first detectionPublication . Cordeiro, Rita; Batista, Fernando da Conceição; Pelerito, Ana; De carvalho, Isabel; Lopo, Sílvia; Neves, Raquel; Rocha, Raquel; Palminha, Paula; Borrego, Maria José; Nuncio, MS; Gomes, João PauloAs mpox continues to spread globally, proactive monitoring and preparedness are crucial to minimize impact and enhance response strategies. Using a mathematical model combining a negative binomial distribution with Richards' logistic curve, we reconstructed the hidden phase of mpox transmission in Portugal, offering insights into the timing and dynamics of the initial outbreak. The analysis of 950 PCR-positive and 986 negative cases suggested that symptom onset occurred between March 24 and April 2, 2022, with March 27 identified as the most probable date. This study delineates the likely period of silent circulation of MPXV in Portugal, providing a clearer understanding of early outbreak dynamics and surveillance performance. Possible imperfections in early diagnostic testing and limited awareness of mpox may have contributed to delayed recognition of the outbreak. By demonstrating how retrospective mathematical modelling can estimate undetected transmission periods, our findings highlight the value of such approaches in epidemic reconstruction and underscore the importance of strengthening early surveillance systems to detect undiagnosed transmission of mpox in non-endemic countries.