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INTRODUCTION: Lymphogranuloma venereum (LGV) is a sexually transmitted infection caused by Chlamydia trachomatis genotypes L1-L3. A combination of techniques with high discriminatory capacity such as multilocus sequence typing (MLST) and the analysis of the ompA gene may be useful to determine the greater penetration of certain strains in transmission networks and their relationship with certain tropisms. AIM: The aim of this study was to investigate the molecular epidemiology of LGV isolates from different regions of Spain. METHODS: Genetic characterisation of LGV isolates detected in six hospitals from Spain between 2018 and 2019 was performed. MLST (five variable regions: hctB, CT058, CT144, CT172 and pbpB) and ompA sequence determination were used to study the LGV strains. RESULTS: Most of the 161 LGV isolates (93.8%) were detected in men who have sex with men (MSM). At least 43.5% of the patients presented with HIV coinfection and 53.4% were symptomatic, with proctitis being the most prevalent symptom (73.3%). Most isolates were detected in Barcelona (n=129).The distribution of ompA genovariants was as follows: 56.1% belonged to L2, 24.3% to L2b, 5.4% to L2bV1, 4.7% to L2bV4, 4.1% to L1, 2.7% to L2b/D-Da, 2.0% to L2bV2 and 0.7% to L2bV7. MLST was successfully performed in 81 samples and 9 different sequence types (STs) were detected. The ompA and MLST combination obtained 17 different genetic profiles, with L2-ST53 and L2-ST58 being the most prevalent (29.5% and 14.1%, respectively). L1 genotype strains belonged to ST23 (n=3) and ST2 (n=3). CONCLUSION: LGV infections were mainly found in MSM living with HIV and with proctitis. The joint analysis of ompA and MLST genetic characterisation techniques showed a high discriminatory capacity. Our findings suggest a cocirculation of L2 and L2b ompA genotypes, and with the inclusion of MLST characterisation, the most prevalent profiles were ompA genotype L2-MLST ST53 and L2-MLST ST58.
ABSTRACT
Annual influenza vaccination is the main strategy to reduce the burden of seasonal influenza epidemics and is recommended for the elderly in most countries with influenza vaccination strategies, with the main objective of preventing hospitalizations and mortality associated with seasonal influenza in this age group. Studies from different countries have estimated the benefits of seasonal influenza vaccination programs in the elderly, preventing a considerable number of cases, hospitalizations and deaths every year. A study measured the number of medically attended confirmed influenza cases in primary care that are prevented annually by vaccination in the population aged 65 and older in Spain, the Netherlands and Portugal, but estimates of the impact of the national influenza vaccination program in the prevention of severe disease in Spain are lacking. The two objectives of this study were to estimate the burden of severe influenza disease in the Spanish population and to measure the impact of influenza vaccination in the prevention of these outcomes in the population aged 65 years and older. Using influenza surveillance systems put in place before the COVID-19 pandemic, we conducted a retrospective observational study to estimate the burden of hospitalizations and ICU admissions in Spain between 2017-18 and 2019-20, by season and age group. Burden estimates for the 65+ group, combined with vaccine effectiveness (VE) and vaccination coverage (VC) data, were used as input data in an ecological, observational study to estimate the impact of the influenza vaccination program on the elderly. We found a higher burden of severe influenza disease in seasons 2017-18 and 2018-19, with A(H3N2) circulation, and in the youngest and oldest age groups. In those aged 65 and older, we estimated an average of 9900 influenza hospitalizations and 1541 ICU admissions averted by vaccination each year. Seasonal influenza vaccination was able to prevent between 11 and 26% influenza hospitalizations and around 40% ICU admissions in the elderly in the three pre-pandemic seasons. In conclusion, our study complements previous analyses in the primary care setting in Spain and demonstrates the benefits of the annual influenza vaccination program in the prevention of severe influenza disease in the elderly, even in seasons with moderate VE.
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The aim of this multicentre project (seven hospitals across the Spanish National Health Service) was to study the phenotypic and genotypic susceptibility of C. trachomatis to the main antimicrobials used (macrolides, doxycycline, and quinolones) in isolates from patients with clinical treatment failure in whom reinfection had been ruled out. During 2018-2019, 73 clinical isolates were selected. Sixty-nine clinical specimens were inoculated onto confluent McCoy cell monolayers for phenotypic susceptibility testing. The minimum inhibitory concentration for azithromycin and doxycycline was defined as the lowest concentration associated with an at least 95% reduction in inclusion-forming units after one passage in the presence of the antibiotic compared to the initial inoculum for each strain (control). Sequencing analysis was performed for the genotypic detection of resistance to macrolides, analysing mutations in the 23S rRNA gene (at positions 2057, 2058, 2059, and 2611), and quinolones, analysing a fragment of the gyrA gene, and searching for the G248T mutation (Ser83->Ile). For tetracyclines, in-house RT-PCR was used to test for the tet(C) gene. The phenotypic susceptibility testing was successful for 10 isolates. All the isolates had minimum inhibitory concentrations for azithromycin ≤ 0.125 mg/L and for doxycycline ≤ 0.064 mg/L and were considered sensitive. Of the 73 strains studied, no mutations were found at positions T2611C or G248T of the gyrA gene. We successfully sequenced 66 isolates. No macrolide resistance-associated mutations were found at positions 2057, 2058, 2059, or T2611C. None of the isolates carried the tet(C) gene. We found no evidence for genomic resistance in this large, clinically relevant dataset.
ABSTRACT
In an increasingly globalized and interconnected world, the outbreak of an infectious disease in one country can become a worrying health emergency for the whole world. A current example is the 2022 monkeypox virus (mpox) outbreak affecting multiple areas across the world. In this context, strategies to interrupt transmission as soon as possible by identifying cases, clusters, and sources of infection should be developed around the world to prevent these crises. The aim of this retrospective and collaborative study was to perform external clinical validation of the VIASURE monkeypox virus real-time PCR detection kit (CerTest Biotec, Spain) with ready-to-use reagents designed for the rapid detection of mpox. A total of 165 samples with suspected infection were used for this analysis. The standard procedures of the clinical microbiology laboratory of the Miguel Servet University Hospital, using the RealStar Orthopoxvirus PCR kit v1.0 (Altona Diagnostics) and bidirectional Sanger sequencing (STAB VIDA, Caparica, Portugal), were considered reference techniques. Furthermore, a subset of 67 mpox-negative samples and 13 mpox-positive samples were routinely tested for clinical diagnosis of other rash/ulcerative pathologies. Accuracy testing resulted in appropriate clinical validation values, as follows: sensitivity, 1 (95% confidence interval [CI], 0.97 to 1); specificity, 1 (95% CI, 0.98 to 1); positive predictive value, 1 (95% CI, 0.93 to 1); negative predictive value, 1 (95% CI, 0.95 to 1). The strength of agreement between assays was almost perfect. The added value is the useful support for the specific diagnosis of mpox infections due to the diagnostic specificity data obtained. IMPORTANCE Given that a large number of mpox outbreaks have been reported worldwide since 2022 in countries in which the disease is not endemic, the main concern for clinicians and global health systems should be to develop effective, available, and easy-to-implement diagnostic strategies to interrupt mpox transmission as soon as possible. This retrospective study demonstrates the satisfactory clinical parameters of a commercially available molecular diagnostic kit for routine testing for mpox in clinical diagnostic laboratories.
Subject(s)
Mpox (monkeypox) , Humans , Mpox (monkeypox)/diagnosis , Mpox (monkeypox)/epidemiology , Real-Time Polymerase Chain Reaction , Retrospective Studies , Disease Outbreaks , LaboratoriesABSTRACT
Chlamydia trachomatis infection is an important public health problem. Our objective was to assess the dynamics of the transmission of this infection, analysing the distribution of circulating ompA genotypes and multilocus sequence types of C. trachomatis in Spain as a function of clinical and epidemiological variables. During 2018 and 2019, we genetically characterized C. trachomatis in tertiary hospitals in six areas in Spain (Asturias, Barcelona, Gipuzkoa, Mallorca, Seville and Zaragoza), with a catchment population of 3.050 million people. Genotypes and sequence types were obtained using polymerase chain reaction techniques that amplify a fragment of the ompA gene, and five highly variable genes (hctB, CT058, CT144, CT172 and pbpB), respectively. Amplicons were sequenced and phylogenetic analysis was conducted. We obtained genotypes in 636/698 cases (91.1%). Overall and by area, genotype E was the most common (35%). Stratifying by sex, genotypes D and G were more common among men, and genotypes F and I among women (p < 0.05). Genotypes D, G and J were more common in men who have sex with men (MSM) than in men who have sex with women (MSW), in whom the most common genotypes were E and F. The diversity index was higher in sequence typing (0.981) than in genotyping (0.791), and the most common sequence types were ST52 and ST108 in MSM, and ST30, ST148, ST276 and ST327 in MSW. Differences in genotype distribution between geographical areas were attributable to differences in population characteristics. The transmission dynamics varied with sexual behaviour: the predominant genotypes and most frequent sequence types found in MSM were different to those detected in MSW and women.
Subject(s)
Chlamydia Infections , Sexual and Gender Minorities , Male , Humans , Female , Homosexuality, Male , Chlamydia trachomatis/genetics , Phylogeny , Multilocus Sequence Typing , Spain/epidemiology , Chlamydia Infections/epidemiology , Genotype , Bacterial Outer Membrane Proteins/geneticsABSTRACT
The capacity of Mycoplasma genitalium to develop resistance to macrolides makes detection of macrolide resistance genes by rapid real-time PCR assays increasingly necessary in clinical diagnostic laboratories so as to initiate appropriate treatment as rapidly as possible. The aim of this retrospective and comparative study was to conduct the clinical evaluation of three commercially available kits for macrolide resistance detection. A total of 111 M. genitalium positive samples analyzed in the Clinical Microbiology Laboratory of the Miguel Servet University Hospital, Zaragoza (Spain) were used. After M. genitalium molecular confirmation, the three assays under study were evaluated and discrepant results were resolved via sequencing. The clinical sensitivity for resistance detection was 83% (95% confidence interval, 69% to 93%) for the ResistancePlus® MG panel kit (SpeeDx Pty Ltd., Sydney, Australia), 95% (84% to 99%) for AllplexTM MG & AziR Assay (Seegene®, Seoul, Korea), and 97% (88% to 99%) for the VIASURE macrolide resistance-associated mutations (23SrRNA) Real time PCR detection kit (Certest Biotec, Zaragoza, Spain). The clinical specificity was 100% (94% to 100%) for Allplex and VIASURE assays and 95% (86% to 99%) for SpeeDx assay. The results arising from this study are cause for strong consideration for the implementation of rapid real-time PCR assays in clinical diagnosis laboratories to eliminate treatment failure and transmission as soon as possible.
Subject(s)
Mycoplasma Infections , Mycoplasma genitalium , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Macrolides/pharmacology , Macrolides/therapeutic use , Mycoplasma genitalium/genetics , Retrospective Studies , Drug Resistance, Bacterial/genetics , Mutation , Mycoplasma Infections/diagnosis , Mycoplasma Infections/drug therapy , Mycoplasma Infections/microbiologyABSTRACT
Introduction. Mycoplasma genitalium is an emerging cause of sexually transmitted infections (STIs) and has been implicated in non-gonococcal urethritis in men and cervicitis in woman. The aim of this study is determinate the incidence and pathogenicity of M. genitalium within the diagnosis of STIs detected from clinical samples in a third level hospital. Material and methods. A total of 8,473 samples from endocervix, urethra, vagina, rectum and others were processed applying Allpex STI Essential Assay. More than 190 records were reviewed to determinate M. genitalium pathogenicity. Results. M. genitalium was detected in a rate 2.8%. Co-infections were detected in 20% of the patients. Conclusions. M. genitalium is considered a STI emerging pathogen thanks to the renewal of multiplex-PCR tests although with a low incidence in our approach. Emerging from our experience and the institutional recommendations both detection of acid nucleic techniques (NAATs) and gonococcal culture might be implemented accurately and coexist to adequate prescriptions. (AU)
Introducción. Mycoplasma genitalium es un patógeno emergente causante de infecciones de transmisión sexual (ITS) y se ha relacionado con uretritis no gonocócica en hombres y cervicitis en mujeres. El objetivo de este estudio es determinar la incidencia y patogenicidad de M. genitalium en el seno del diagnóstico de ITS detectadas a partir de muestras clínicas en un hospital terciario. Métodos. Se procesaron 8.473 muestras de endocérvix, uretra, vagina, recto y otros, aplicando Allpex STI Essential Assay. Se revisaron más de 190 historias clínicas para determinar la patogenicidad de M. genitalium. Resultados. Se detectó M. genitalium en 2,8% de casos. Hubo coinfecciones en 20% de los pacientes. Conclusiones. M. genitalium a pesar de la baja incidencia en nuestra revisión, actualmente es un patógeno de valor en alza gracias al desarrollo de técnicas moleculares como PCRmultiplex. A partir de nuestra experiencia y las recomendaciones institucionales, tanto las técnicas de detección de ácidos nucleicos (NAATs) como los cultivos para gonococo deberían implementarse y coexistir para adecuar los tratamientos (AU)
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Humans , Mycoplasma genitalium , Sexually Transmitted Diseases , Primary Health Care , Spain , Urethritis , Uterine CervicitisABSTRACT
No disponible
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Humans , Male , Aged , Mycetoma/diagnosis , Hemoptysis/etiology , Lung Diseases, Fungal/diagnosis , Embolization, Therapeutic , PneumonectomyABSTRACT
No disponible
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Humans , Female , Young Adult , Tinea Capitis/diagnosis , Microsporidia/isolation & purification , Vulvar Diseases/microbiologyABSTRACT
No disponible
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Adolescent , Female , Humans , Pneumonia, Bacterial , Legionellosis , LegionellaABSTRACT
No disponible