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Percorrer por autor "Roque, Carla"

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  • An Overview of Monkeypox Virus Detection in Different Clinical Samples and Analysis of Temporal Viral Load Dynamics
    Publication . Cordeiro, Rita; Pelerito, Ana; de Carvalho, Isabel Lopes; Lopo, Sílvia; Neves, Raquel; Rocha, Raquel; Palminha, Paula; Verdasca, Nuno; Palhinhas, Cláudia; Borrego, Maria José; Manita, Carla; Ferreira, Idalina; Bettencourt, Célia; Vieira, Patrícia; Silva, Sónia; Água-Doce, Ivone; Roque, Carla; Cordeiro, Dora; Brondani, Greice; Santos, João Almeida; Martins, Susana; Rodrigues, Irene; Ribeiro, Carlos; Núncio, Maria Sofia; Gomes, João Paulo; Batista, Fernando da Conceição
    Mpox is a zoonotic disease caused by the Monkeypox virus (MPXV), and since May 2022, tens of thousands of cases have been reported in non-endemic countries. We aimed to evaluate the suitability of different sample types for mpox diagnostic and assess the temporal dynamics of viral load. We evaluated 1914 samples from 953 laboratory-confirmed cases. The positivity rate was higher for lesion (91.3%) and rectal swabs (86.1%) when compared with oropharyngeal swabs (69.5%) and urines (41.2%), indicating higher viral loads for the former. Supporting this, lesion and rectal swabs showed lower median PCR C values (C = 23 and C = 24), compared to oropharyngeal swabs and urines (C = 31). Stable MPXV loads were observed in swabs from lesions up to 30 days after symptoms onset, contrasting with a considerable decrease in viral load in rectal and oropharyngeal swabs. Overall, these results point to lesion swabs as the most suitable samples for detecting MPXV in the 2022-2023 multicountry outbreak and show comparable accuracy to rectal swabs up to 8 days after symptoms onset. These findings, together with the observation that about 5% of patients were diagnosed through oropharyngeal swabs while having negative lesions, suggest that multisite testing should be performed to increase diagnostic sensitivity.
    2024-12Artigo de investigação Acesso aberto Ver mais
  • Caraterização virológica dos vírus da gripe que circularam em Portugal na época 2014/2015
    Publication . Pechirra, Pedro; Costa, Inês; Cristóvão, Paula; Roque, Carla; Barreiro, Paula; Duarte, Sílvia; Machado, Ausenda; Rodrigues, Ana Paula; Nunes, Baltazar; Guiomar, Raquel
    Introdução e objetivo: A monitorização contínua das propriedades antigénicas e genéticas dos vírus da gripe é essencial, quer para a seleção anual das estirpes virais a incluir na vacina, quer para identificar novas linhas de orientação da terapêutica antiviral. O presente estudo descreve as caraterísticas antigénicas e genéticas dos vírus da gripe identificados em Portugal no inverno de 2014/2015.
    2015-11Artigo científico Acesso aberto Ver mais
  • Diagnóstico laboratorial do sarampo em Portugal, 2011-2013
    Publication . Palminha, Paula; Vinagre, Elsa; Cordeiro, Rita; Ribeiro, Carlos; Roque, Carla
    Objetivo: Este estudo tem como objetivo descrever os casos prováveis de sarampo enviados ao INSA para confirmação laboratorial entre 2011 e 2013 em Portugal.
    2015-12-22Artigo científico Acesso aberto Ver mais
  • Enterovirus D68 diagnosed in severe respiratory and neurological illness in children during 2015-2016 season in Portugal
    Publication . Guiomar, Raquel; Costa, Inês; Pechirra, Pedro; Palminha, Paula; Ribeiro, Carlos; Roque, Carla; Peres, Maria João; Viseu, Regina; Balseiro, Maria Jesus; Brito, Maria João; Neves, João; Branquinho, Paula; Côrte-Real, Rita
    Background: Enterovirus D68 (EV-D68) was first isolated in 1962, and since then associated with respiratory illness. The report of severe respiratory and neurological disease including deaths associated to EV-D68 in United States and Canada during August 2014 highlighted the need of epidemiological information regarding EV-D68 circulation. In Europe information was scarce, available only for few countries. In Portugal there was no data available and was critical to know the epidemiology of EV-D68, especially in children hospitalized with severe respiratory or neurological disease. This study aims to identify EV-D68 in Enterovirus positive respiratory samples in children under 18 with clinical diagnosis of severe respiratory infection or neurological illness. Methods: During 2015/16 winter season, between November/2015 and March/2016, 29 EV positive cases were reported to the National Influenza and Other Respiratory Virus Reference Laboratory (NIC) by two hospitals located in Lisbon and Setubal districts. EV diagnosis was performed in hospitals by biomolecular methods using commercial kits (real time multiplex-PCR, FTD Respiratory pathogens 21 and CLART Pneumovir, Genomica, respectively). EV-D68 was diagnosed by an in house real-time PCR [1]. Virus isolation in RD cell line and phylogenentic analysis of the VP1/VP3 genomic regions will enable the identification of genetic groups in circulation. All samples were irreversibly anonymized. Demographic and clinical data were collected. Results: EV-D68 was confirmed in 20 respiratory samples previously positive for EV (69%; 20/29). Samples were collected from children with age ranging from 2 months to 6 years old, both genders (9 female; 11 male) with diagnosis of severe respiratory or neurological illness. Eighteen cases were hospitalized (90%; 18/20). Bronchiolitis and pneumonia were the most frequently reported diagnosis, corresponding to 70% (14/20). Two cases have neurologic diagnosis. EV-D68 was identified throughout all study period with the higher number of positive cases detected during January 2016, in week 3. Virus isolation and genetic characterization are under way with expected results in virus phylogeny and evaluation on similarity with recent circulating strains in United States, Canada and European countries. Conclusions: EV-D68 was detected in a high positive rate (69%) among EV positive cases. This positive rate of EV-D68 was higher compared to the positivity rate of 10,2 %, calculated in a European study during 2014 [2]. This finding could be linked to the selection of severe and hospitalized patients in present study, highlighting the involvement of EV-D68 with severe respiratory disease in children. The identification of EV-D68 is also crucial in respiratory samples in children with clinical diagnosis of neurological illness. This study is the first attempt to describe the prevalence of EV-D68 in severe paediatric cases, in Portugal. The strength of EV-D68 surveillance in paediatric and adult population at the national level will be important to understand the epidemiology of EV-D68, age-related susceptibility and association with disease severity.
    2016-09-14Documento de conferência Acesso aberto Ver mais
  • Estudo da imunidade adquirida pela vacinação contra a COVID-19 em profissionais de saúde do INSA: relatório de execução
    Publication . Guiomar, Raquel; Santos, Ana João; Melo, Aryse; Ramalhete, Sara; Costa, Inês; Henriques, Camila; Matos, Rita; Rodrigues, Ana Paula; Irina, Kislaya; Silva, Anabela Santos; Roque, Carla; Silva, Carla; Aguiar, Joaquim; Dias, Carlos; Machado, Jorge; Graça, Fátima; Graça, António Silva; Machado, Ausenda
    O presente relatório de execução, elaborado pelo Departamento de Epidemiologia, o Departamento de Doenças Infeciosas e os Serviços de Saúde Ocupacional do INSA, apresenta os resultados de implementação dos 12 meses do estudo, desenvolvido para estimar a efetividade da vacina contra a COVID-19 em profissionais de saúde do Instituto Nacional de Saúde Doutor Ricardo Jorge (INSA). Neste relatório é apresentada informação sobre a execução do estudo, incluindo descrição dos participantes e da sua participação nos diferentes momentos e etapas, bem como a informação recolhida através das diferentes fontes de dados. O estudo prospetivo de coorte teve como principal objetivo seguir grupos de profissionais vacinados e não-vacinados durante 12 meses para estimar a efetividade da vacina contra a COVID-19 nos trabalhadores do INSA e analisar a resposta imunológica humoral através da deteção de anticorpos vacinais. O estudo iniciou-se em janeiro de 2021 e terminou a 16 fevereiro 2022.
    2022-05Relatório Acesso restrito Ver mais
  • Genital Herpes in a STD outpatient clinic in Lisbon
    Publication . Lopo, Sílvia; Roque, Carla; Costa, Inês; Borrego, Maria José; Azevedo, Jacinta
    Introduction: Genital Herpes is the major cause of genito-ulcerative disease affecting a considerable number of individuals worldwide and is a chronic, life-long viral infection caused by both HSV1 or HSV2. Most cases of recurrent genital herpes are caused by HSV2, being the leading cause of genital ulcer disease in developing countries, but the proportion of anogenital herpetic infections attributed to HSV1 are increasing, especially in young women and MSM. The prevalence in the general population ranges from 10% to 80% and depends on socio-economic factors. Seropositivity rates are higher in women than in men and increase with age. Reactivation and subclinical shedding is more frequent in genital infections caused by HSV2 than by HSV1, which reaffirm the importance of laboratory confirmation of clinical diagnosis. Aims: Retrospective study of the role of HSV1 and HSV2 infections in genital ulcerations from a population of a Sexually Transmitted Diseases Outpatient Clinic, according to epidemiological, laboratory and clinical data. Methodology: 56 ulcer genital/urethral swabs from patients suspected of HSV infection were sent to the National Institute of Health (NIH), in Lisbon, between April 2015–April 2016. HSV1 and HSV2 were determined by a quantitative commercial real-time PCR kit, which targets a fragment of 162 bp of a region located in the US7 gene for HSV1 and a fragment of 177 bp of a region located in the US2 gene for HSV2. The 56 swabs were also inoculated in Vero cell cultures for determination of cytopathic effect. Results: HSV infection in genital/urethral swabs were detected in 30 (53.6%) of 56 samples. The symptoms of the positive cases were genital ulcerations in vulva or penis and/or perineum and the clinical diagnosis was genital herpes infection. In 7 of the 30 positive cases (23.3%) HSV1 DNA was detected (2 man and 5 women with age ranges between 17 and 27 years old); and in 23 of the 30 positive cases (76.7%) HSV2 DNA was detected (18 man and 5 women, with age ranges between 17 and 62 years old). Five of the 7 HSV1 positive cases were primoinfections (71.4%) and in the 23 HSV2 positive cases, 3 (13.0%) were primoinfections and 8 (34.8%) were the first ulcer episode but not primoinfections. HSV DNA viral load values varied between 21848–87474493 cop/ml in HSV1 cases and between 1177– 31160846336 cop/ml in HSV2 cases. We didn’t find direct correlation between viral load and primary vs recurrent infection although the higher viral load was found in HSV2 first episode cases. Cytophatic effect was observed in all positive PCR cases. All positive cases were treated with valacyclovir and resolved after treatment. Comments: To identify HSV genital infections is important for the specific treatment, for preventing the transmission of HSV to partners, and to prevent the risk of acquiring and transmitting HIV. In our study 53.6% cases were positive for HSV genital infection; because of social, demographic and migratory tendency, the population at risk for STI continues to grow and experience an increased burden of disease. We also observed in this population an increasing proportion of HSV1 genital primoinfection, which is in accordance to the literature. In the present study we confirmed the usefulness of real-time PCR for HSV DNA detection in genital ulcerations. Concerning the correlation of viral load with subtype, the differences should be further evaluated with an increase number of clinical cases.
    2016-09Documento de conferência Acesso restrito Ver mais
  • A gripe em Portugal: análise preliminar da atividade gripal 2014/2015
    Publication . Pechirra, Pedro; Cristóvão, Paula; Costa, Inês; Roque, Carla; Barreiro, Paula; Duarte, Sílvia; Machado, Ausenda; Rodrigues, Ana Paula; Nunes, Baltazar; Guiomar, Raquel
    Objetivo: Pretende-se com a presente publicação, divulgar a análise preliminar da atividade gripal em Portugal na época de 2014/2015 (entre 15 de setembro de 2014 e 20 de março de 2015).
    2015-05Artigo científico Acesso aberto Ver mais
  • Influenza in Portugal, 2014/2015 season
    Publication . Pechirra, Pedro; Cristóvão, Paula; Costa, Inês; Roque, Carla; Barreiro, Paula; Duarte, Sílvia; Machado, Ausenda; Rodrigues, Ana Paula; Nunes, Baltazar; Guiomar, Raquel
    During 2014/2015 season the influenza activity was high, the epidemic period was observed between week 1/2015 and 8/2015 with a maximum of 148 ILI cases per 105 inhabitants in week 4/2015. The influenza B virus, A(H3) and A(H1)pdm09 co-circulated during the season, although influenza B were dominant throughout the winter, except for weeks 4, 6 and 7/2015 where it was detected in co-dominance with influenza A virus, situation that contrasts with European influenza picture. The antigenic and genetic analysis of circulating influenza A(H3) and B virus showed differences regarding 2014/2015 influenza vaccine strains. 70% of A(H3) viruses belonged to subgroup 3C.2a, antigenically different from vaccine strain. A (H3) positive were recorded in higher percentage in children (0 to 4 years: 40.0% and 5 to 14 years: 39.7%) and in adults over 65 years (36, 8%). A(H1)pdm09 virus were sporadically detected and were similar to the vaccine A/California/7/2009 strain. In general, the detected influenza viruses were similar to those recommended strains for next winter`s vaccine 2015/2016.
    2015-06-12Documento de conferência Acesso aberto Ver mais
  • Investigating the impact of COVID-19 vaccines on the red blood cell immune function by omics-based approaches
    Publication . Saraiva, Joana; Coelho, Cristina Valentim; Vaz, Fátima; Antunes, Marilia; Neves, Sofia; Ricardo, Peliano; Andrade, Odília; Miranda, Armandina; Melo, Aryse; Roque, Carla; Guiomar, Raquel; Mohammad, Hamza; Soares, Nelson; Penque, Deborah
    The role of red blood cells (RBC) in the immune system is increasingly recognized. However, RBC-derived molecules with an immunomodulatory role in health and disease, as well as in vaccine immunogenicity are still poorly investigated. Taking as a model the emergent COVID-19 vaccines, we aimed to investigate whether vaccines induce proteome and/or metabolome changes in RBCs able to affect T-cell immune activity, as a mechanistic test for vaccine immunization regulated by RBCs. Our ultimate goal is to identify RBC immunomodulators as potential co-adjuvants in the formulation of next-generation vaccines with bolstered efficacy and duration. A biobank of blood samples collected longitudinally under ‘omics’ quality control from subjects (n=39) that underwent vaccination for COVID-19 between April and September 2021 was created. This biobank is associated with extensive clinical data, including demographic data, COVID-19 PCR diagnosis, hematological and vaccine effectivity data. Linear Mixed Models, were used to evaluate the association between biometrical characteristics, health related habits, vaccine technology and vaccine effectivity and hematological parameters, along the different time-points (t0-t4) under study, i.e, before and after (24-72h or 30 days) of the first and second dose of vaccine. Statistical analyses were performed using R software version 4.1.2. Results showed significant differences (p<0.05) before/after vaccination in a set of hematological variables (e.g., hemoglobin, lymphocytes and monocytes values), as well in terms of vaccine effectivity and vaccine technology (mRNA or adenovirus – based vaccines). Preliminary data from proteomics and metabolomics analysis of RBCs along the different time-points (t0-4) of immunization response will be also presented and discussed. The knowledge gained with this project can generate important evidence-based recommendations intended to optimize vaccine immunization, by recognizing the impact of blood cells such RBCs in the immune system regulation.
    2022-05Documento de conferência Acesso aberto Ver mais
  • Molecular studies on HSV: replication rate, infection capacity and progeny
    Publication . Azevedo, Aleksandra; Nunes, Alexandra; Roque, Carla; Costa, Inês; Gomes, João Paulo; Lopo, Sílvia
    Introduction: In the last years genital herpes has emerged as one of the most prevalent sexually transmitted infections. Herpes simplex virus (HSV) is the most common cause of genital ulcer disease, with infections caused by both sub-types HSV-1 and HSV-2. A better understanding of the virus replication cycle is relevant to the pathogenesis of human diseases and is essential for the development of antiviral chemotherapy. Objectives: We aimed to shed some light on the HSV-1 and HSV-2 infectious cycle, namely their capacity of infection, replication rate and progeny, in three distinct cell lines (Vero, Vero E6 and HeLa229). We also aimed to evaluate whether the concentration of virus has any influence on the degree of the infection. Methodology: Preliminary assays were performed in order to understand which cellular concentration, viral load, nutrients’ availability and inoculation modus operandi (centrifugation versus agitation) best mimic the HSV infection. Confluent cell monolayers were infected with two HSV-2 and two HSV-1 at MOIs of 1:10, 1:1, 10:1 and 100:1. Inoculations were performed in parallel in two 24-well plates, one for quantitative real-time PCR (kPCR) and one for immunofluorescence assays, which were incubated for 30 hours at 37ºC and 5% CO2. At different times-points of infection (6, 12, 18, 24 and 30 hours p.i.), the wells were scratched for kPCR and the slides were stained with monoclonal antibodies. For kPCR assays, appropriate standard curves were generated by serial diluting plasmids cloned with HSV-1 and HSV-2 single copy genes. Results and Conclusions: Preliminary assays showed that, regardless of the viral load, it takes approximately 23 hours for the virus to complete the infectious cycle taking into account that no replication is observed after this time point. Considering the comparison between the two inoculation procedures (centrifugation versus agitation), we only observed relevant differences for lower viral loads, with centrifugation yielding more viral progeny. More specific data regarding both the HSV-1 and HSV-2 replication capacity for different MOIs are currently under evaluation.
    2016-09Documento de conferência Acesso restrito Ver mais