Browsing by Author "Azevedo, Jacinta"
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- Carga treponémica em amostras biológicas correspondentes a diferentes fases clínicas de sífilisPublication . Pinto, Miguel; Antelo, Minia; Ferreira, Rita; Azevedo, Jacinta; Santo, Irene; Borrego, Maria José; Gomes, João Paulo[PT] Sífilis, a doença sexualmente transmitida causada por Treponema pallidum, apresenta diferentes manifestações clinicas. No entanto, os fatores que levam às diversas progressões da doença permanecem desconhecidos. Este estudo teve como objetivo a quantificação da carga treponémica, em diversas amostras biológicas, para contribuir para uma melhor compreensão da doença. Analisaram-se 309 amostras de DNA de distintos locais anatómicos associados a diferentes manifestações de sífilis. Uma quantificação absoluta por PCR em tempo-real foi utilizada para quantificar precisamente o número de células treponémicas e humanas, e calcular a carga treponémica em cada amostra. Os exsudados de lesões primárias apresentaram as cargas treponémicas mais elevadas contrastando com as amostras sanguíneas. Uma grande dispersão de concentrações bacterianas foi observada nas amostras sanguíneas dos casos de sífilis secundária. T. pallidum foi detetado em 37 amostras de indivíduos seronegativos e em 13 casos de sífilis tratada. Este estudo sugere um cenário onde a sífilis poderá ser caracterizada por: i) cargas elevadas e heterogéneas na sífilis primária, refletindo a duração do desenvolvimento e resolução da lesão; ii) uma potencial capacidade replicativa de T. pallidum na corrente sanguínea, sugerida pela elevada dispersão de concentrações na sífilis secundária; e iii) uma evasão bacteriana quer ao sistema imunitário do hospedeiro quer ao tratamento antibiótico.
- Chlamydia trachomatis outbreak: when the virulence-associated genome backbone imports a prevalence-associated major antigen signaturePublication . Borges, Vitor; Cordeiro, Dora; Salas, Ana Isabel; Lodhia, Zohra; Correia, Cristina; Isidro, Joana; Fernandes, Cândida; Rodrigues, Ana; Azevedo, Jacinta; Alves, João; Rôxo, João; Rocha, Miguel; Corte-Real, Rita; Vieira, Luís; Borrego, Maria José; Gomes, João PauloChlamydia trachomatis is the most prevalent sexually transmitted bacterium worldwide and the causative agent of trachoma. Its strains are classified according to their ompA genotypes, which are strongly linked to differential tissue tropism and disease outcomes [ocular disease, urogenital disease and lymphogranuloma venereum (LGV)]. While the genome-based species phylogenetic tree presents four main clades correlating with tropism/prevalence, namely ocular, LGV, urogenital T1 (more prevalent genotypes) and urogenital T2 (less prevalent genotypes), inter-clade exchange of ompA is considered a rare phenomenon probably mediating marked tropism alterations. An LGV epidemic, associated with the clonal expansion of the L2b genotype, has emerged in the last few decades, raising concerns particularly due to its atypical clinical presentation (ulcerative proctitis) and circulation among men who have sex with men (MSM).
- Chlamydia trachomatis: when the virulence-associated genome backbone imports a prevalence-associated major antigen signaturePublication . Borges, Vítor; Cordeiro, Dora; Salas, Ana Isabel; Lodhia, Zohra; Correia, Cristina; Isidro, Joana; Fernandes, Cândida; Rodrigues, Ana Maria; Azevedo, Jacinta; Alves, João; Roxo, João; Rocha, Miguel; Côrte-Real, Rita; Vieira, Luís; Borrego, Maria José; Gomes, João PauloChlamydia trachomatis is the most prevalent sexually transmitted bacterium worldwide and the causative agent of trachoma. Its strains are classified according to their ompA genotypes, which are strongly linked to differential tissue tropism and disease outcomes [ocular disease, urogenital disease and lymphogranuloma venereum (LGV)]. While the genome-based species phylogenetic tree presents four main clades correlating with tropism/prevalence, namely ocular, LGV, urogenital T1 (more prevalent genotypes) and urogenital T2 (less prevalent genotypes), inter-clade exchange of ompA is considered a rare phenomenon probably mediating marked tropism alterations. An LGV epidemic, associated with the clonal expansion of the L2b genotype, has emerged in the last few decades, raising concerns particularly due to its atypical clinical presentation (ulcerative proctitis) and circulation among men who have sex with men (MSM). Here, we report an LGV outbreak, mostly affecting human immunodeficiency virus-positive MSM engaging in high-risk sexual practices, caused by an L2b strain with a rather unique non-LGV ompA signature that precluded the laboratory notification of this outbreak as LGV. C. trachomatis whole-genome capture and sequencing directly from clinical samples was applied to deeply characterize the genomic backbone of this novel LGV outbreak-causing clone. It revealed a chimeric genome structure due to the genetic transfer of ompA and four neighbouring genes from a serovar D/Da strain, likely possessing the genomic backbone associated with the more prevalent urogenital genotypes (T1 clade), to an LGV (L2b) strain. The hybrid L2b/D-Da strain presents the adhesin and immunodominant antigen MOMP (major outer membrane protein) (encoded by ompA) with an epitope repertoire typical of non-invasive genital strains, while keeping the genome-dispersed virulence fingerprint of a classical LGV strain. As previously reported for inter-clade ompA exchange among non-LGV clades, this novel C. trachomatis genomic mosaic involving a contemporary epidemiologically and clinically relevant LGV strain may have implications on its transmission, tissue tropism and pathogenic capabilities. The emergence of variants with epidemic and pathogenic potential highlights the need for more focused surveillance strategies to capture C. trachomatis evolution in action.
- Clinical and Epidemiological Characterization of Lymphogranuloma Venereum in a Sexually Transmitted Diseases Clinic in Lisbon, 2001 to 2020Publication . Bonito, Frederico; Alves, João; Lodhia, Zohra; Cordeiro, Dora; Borges, Vítor; Azevedo, Jacinta; Borrego, Maria JoséBetween 2001 and 2020, 54 LGV cases were diagnosed in a sexually transmitted disease clinic in Lisbon, most in men who have sex with men (87%), HIV negative (63%), from the anorectal mucosa (72.2%). Cases among heterosexuals were also identified (13%). Surveillance programs irrespective of sexual orientation and HIV status are needed to avoid the morbidity associated with LGV.
- Diagnóstico laboratorial da infeção por Chlamydia trachomatis, 1991-2014Publication . Dinis, Margarida; Cordeiro, Dora; Santo, Irene; Azevedo, Jacinta; Gomes, João Paulo; Borrego, Maria JoséObjetivo: O presente trabalho tem por objetivo apresentar os resultados do diagnóstico laboratorial das infeções por C. trachomatis realizado no Instituto Nacional de Saúde Doutor Ricardo Jorge (INSA) entre 1991 e 2014.
- Europe-wide expansion and eradication of multidrug-resistant Neisseria gonorrhoeae lineages: a genomic surveillance studyPublication . Sánchez-Busó, Leonor; Cole, Michelle J.; Spiteri, Gianfranco; Day, Michaela; Jacobsson, Susanne; Golparian, Daniel; Sajedi, Noshin; Yeats, Corin A.; Abudahab, Khalil; Underwood, Anthony; Bluemel, Benjamin; Aanensen, David M.; Unemo, Magnus; Pleininger, Sonja; Indra, Alexander; De Baetselier, Irith; Vanden Berghe, Wim; Hunjak, Blaženka; Blažić, Tatjana Nemeth; Maikanti-Charalambous, Panayiota; Pieridou, Despo; Zákoucká, Hana; Žemličková, Helena; Hoffmann, Steen; Cowan, Susan; Schwartz, Lasse Jessen; Peetso, Rita; Epstein, Jevgenia; Viktorova, Jelena; Ndeikoundam, Ndeindo; Bercot, Beatrice; Bébéar, Cécile; Lot, Florence; Buder, Susanne; Jansen, Klaus; Miriagou, Vivi; Rigakos, Georgios; Raftopoulos, Vasilios; Balla, Eszter; Dudás, Mária; Ásmundsdóttir, Lena Rós; Sigmundsdóttir, Guðrún; Hauksdóttir, Guðrún Svanborg; Gudnason, Thorolfur; Colgan, Aoife; Crowley, Brendan; Saab, Sinéad; Stefanelli, Paola; Carannante, Anna; Parodi, Patrizia; Pakarna, Gatis; Nikiforova, Raina; Bormane, Antra; Dimina, Elina; Perrin, Monique; Abdelrahman, Tamir; Mossong, Joël; Schmit, Jean-Claude; Mühlschlegel, Friedrich; Barbara, Christopher; Mifsud, Francesca; Van Dam, Alje; Van Benthem, Birgit; Visser, Maartje; Linde, Ineke; Kløvstad, Hilde; Caugant, Dominique; Młynarczyk-Bonikowska, Beata; Azevedo, Jacinta; Borrego, Maria-José; Nascimento, Marina Lurdes Ramos; Pavlik, Peter; Klavs, Irena; Murnik, Andreja; Jeverica, Samo; Kustec, Tanja; Vázquez Moreno, Julio; Diaz, Asuncion; Abad, Raquel; Velicko, Inga; Unemo, Magnus; Fifer, Helen; Shepherd, Jill; Patterson, LynseyBackground: Genomic surveillance using quality-assured whole-genome sequencing (WGS) together with epidemiological and antimicrobial resistance (AMR) data is essential to characterise the circulating Neisseria gonorrhoeae lineages and their association to patient groups (defined by demographic and epidemiological factors). In 2013, the European gonococcal population was characterised genomically for the first time. We describe the European gonococcal population in 2018 and identify emerging or vanishing lineages associated with AMR and epidemiological characteristics of patients, to elucidate recent changes in AMR and gonorrhoea epidemiology in Europe. Methods: We did WGS on 2375 gonococcal isolates from 2018 (mainly Sept 1-Nov 30) in 26 EU and EEA countries. Molecular typing and AMR determinants were extracted from quality-checked genomic data. Association analyses identified links between genomic lineages, AMR, and epidemiological data. Findings: Azithromycin-resistant N gonorrhoeae (8·0% [191/2375] in 2018) is rising in Europe due to the introduction or emergence and subsequent expansion of a novel N gonorrhoeae multi-antigen sequence typing (NG-MAST) genogroup, G12302 (132 [5·6%] of 2375; N gonorrhoeae sequence typing for antimicrobial resistance [NG-STAR] clonal complex [CC]168/63), carrying a mosaic mtrR promoter and mtrD sequence and found in 24 countries in 2018. CC63 was associated with pharyngeal infections in men who have sex with men. Susceptibility to ceftriaxone and cefixime is increasing, as the resistance-associated lineage, NG-MAST G1407 (51 [2·1%] of 2375), is progressively vanishing since 2009-10. Interpretation: Enhanced gonococcal AMR surveillance is imperative worldwide. WGS, linked to epidemiological and AMR data, is essential to elucidate the dynamics in gonorrhoea epidemiology and gonococcal populations as well as to predict AMR. When feasible, WGS should supplement the national and international AMR surveillance programmes to elucidate AMR changes over time. In the EU and EEA, increasing low-level azithromycin resistance could threaten the recommended ceftriaxone-azithromycin dual therapy, and an evidence-based clinical azithromycin resistance breakpoint is needed. Nevertheless, increasing ceftriaxone susceptibility, declining cefixime resistance, and absence of known resistance mutations for new treatments (zoliflodacin, gepotidacin) are promising.
- Genital Herpes in a STD outpatient clinic in LisbonPublication . Lopo, Sílvia; Roque, Carla; Costa, Inês; Borrego, Maria José; Azevedo, JacintaIntroduction: 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.
- Genome-scale analysis of the non-cultivable Treponema pallidum reveals extensive within-patient genetic variationPublication . Pinto, Miguel; Borges, Vítor; Antelo, Minia; Pinheiro, Miguel; Nunes, Alexandra; Azevedo, Jacinta; Borrego, Maria José; Mendonça, Joana; Carpinteiro, Dina; Vieira, Luís; Gomes, João Paulo.Insights into the genomic adaptive traits of Treponema pallidum, the causative bacterium of syphilis, have long been hampered due to the absence of in vitro culture models and the constraints associated with its propagation in rabbits. Here, we have bypassed the culture bottleneck by means of a targeted strategy never applied to uncultivable bacterial human pathogens to directly capture whole-genome T. pallidum data in the context of human infection. This strategy has unveiled a scenario of discreet T. pallidum interstrain single-nucleotide-polymorphism-based microevolution, contrasting with a rampant within-patient genetic heterogeneity mainly targeting multiple phase-variable loci and a major antigen-coding gene (tprK). TprK demonstrated remarkable variability and redundancy, intra- and interpatient, suggesting ongoing parallel adaptive diversification during human infection. Some bacterial functions (for example, flagella- and chemotaxis-associated) were systematically targeted by both inter- and intrastrain single nucleotide polymorphisms, as well as by ongoing within-patient phase variation events. Finally, patient-derived genomes possess mutations targeting a penicillin-binding protein coding gene (mrcA) that had never been reported, unveiling it as a candidate target to investigate the impact on the susceptibility to penicillin. Our findings decode the major genetic mechanisms by which T. pallidum promotes immune evasion and survival, and demonstrate the exceptional power of characterizing evolving pathogen subpopulations during human infection.
- Nova abordagem genómica para a investigação da sífilisPublication . Pinto, Miguel; Borges, Vítor; Antelo, Minia; Pinheiro, Miguel; Nunes, Alexandra; Azevedo, Jacinta; Borrego, Maria José; Mendonça, Joana; Carpinteiro, Dina; Vieira, Luís; Gomes, João PauloA sífilis é uma doença sexualmente transmissível causada pela bactéria Treponema pallidum e constitui um problema de saúde pública mundial, em parte devido à ausência de uma vacina para prevenção da sua transmissão. A investigação desta doença tem sido atrasada pela incapacidade histórica de cultivar T. pallidum in vitro, dificultando por exemplo o desenvolvimento de estudos genómicos. De facto, há uma grande lacuna no conhecimento da epidemiologia molecular deste agente patogénico, assim como da base molecular que medeia a patogénese da sífilis. No estudo aqui apresentado, foi possível implementar uma abordagem inovadora para capturar o genoma de T. pallidum no contexto de infeção humana, evitando-se, assim, a necessidade da cultura da bactéria em modelo animal. Esta estratégia permitiu estudar, pela primeira vez, como é que este agente patogénico vai alterando o seu genoma para se adaptar e sobreviver como agente infecioso humano. Nomeadamente permitiu descodificar os principais mecanismos genéticos pelos quais a bactéria T. pallidum evade o sistema imunitário e se adapta ao Homem nesta complexa e multifásica doença. A aplicação desta estratégia inovadora de monitorização da interação Homem-bactéria poderá ser importante para o desenvolvimento de novas medidas profiláticas e/ou terapêuticas. Acresce que esta abordagem constitui também um ponto de viragem para o aperfeiçoamento de metodologias de diagnóstico e de epidemiologia molecular, o que permitirá aumentar o conhecimento da distribuição geográfica, das vias de transmissão e das propriedades de virulência deste agente patogénico para bem da saúde pública.
- Prevalence of four urogenital sexually transmitted infections in a dedicated clinic from LisbonPublication . Lodhia, Zohra; Azevedo, Jacinta; Alves, João; Cordeiro, Dora; Antunes, Marília; Borrego, Maria J.Background/Objectives: To determine the prevalence of urogenital Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV), and Mycoplasma genitalium (MG) among attendees of an open and freely available sexually transmitted infections (STI) dedicated clinic in Lisbon, at Centro de Saúde da Lapa, during 1-year. Methods: Molecular testing for CT, NG, MG, and TV was performed on 1,062 urogenital specimens (one specimen per person). A descriptive, cross- sectional, observational study was conducted to evaluate the characteristics of infected persons. Statistical analysis was performed. Results: Around 237 infections were detected in 214 patients. CT was the most prevalent (11.6%), with a similar infection rate between men and women. NG was the second most frequently detected (7.3%), followed by MG and TV (2.9 and 0.5%, respectively). Statistically significant associations were found: 1) between younger age and CT and NG prevalence, where being < 25 years old constituted an increased risk factor; 2) between CT and NG prevalence and sexual orientation, where heterosexuals presented an increased risk for CT infections while men who have sex with men (MSM) had a higher risk for NG infections; and 3) between “having symptoms” and gonococcal infection. Conclusions: This study highlights the rising of CT and NG in contrast to a low rate of MG and to the scarceness of TV.