Objective: Coronavirus disease 2019 (COVID-19) caused increased intensive care unit admissions, invasive procedures, and antimicrobial use, potentially worsening bacterial infections and multidrug resistance. Retrospective studies have found that Klebsiella pneumoniae co-infections in COVID-19 patients were significant and possibly linked to mechanical ventilation and central catheter placement. In the present study, we assessed the genetic diversity and antimicrobial resistance of K. pneumoniae isolates collected before and during the pandemic to evaluate the effect of the pandemic on these factors. Methods: Antimicrobial resistance profiling, wholegenome sequencing, and phylogenetic analyses were used to examine resistance patterns, genetic diversity, and mobile genetic elements. Results: From January 2018 to January 2021, 263 K. pneumoniae isolates were identified from infection sites. Carbapenem-resistant isolates increased from 32.5% in 2019 to 60.3% in 2020, remaining stable until January 2021. Nevertheless, healthcare-associated infections did not increase significantly, highlighting the effectiveness of infection control programs. Whole-genome sequencing showed that 54% of carbapenem-resistant isolates carried plasmids resembling pKPC_FCF3SP (IncN) and pKpQIL-like (IncFII) plasmids; however, the number of pKpQIL-like plasmid carriers declined during the pandemic likely due to patient transfer carrying isolates with distinct mobilome. Simultaneously, the number of plasmidnegative isolates increased by 25%. Carbapenem-resistant isolates showed multidrug resistance, particularly to cephalosporins and fluoroquinolones; however, aminoglycosides remained effective. Genetic analysis identified ten aminoglycoside resistance genes, with aac(6')-Ib-D181Y, associated with improved substrate recognition, being the most prevalent. Virulence factors included four integrative conjugative elements, with the integrative conjugative element K. pneumoniae ICEKp4 found only in pandemic period isolates. The O4 and O2 antigens predominated, whereas O3b appeared exclusively during the pandemic. Conclusion: The COVID-19 pandemic has contributed to a rise in carbapenem-resistant K. pneumoniae, underscoring the need for ongoing surveillance. Molecular shifts reflect the adaptation of the pathogen to evolving clinical settings.

Objective:This study aimed to evaluate the Trueprep/Truelab platform with four Truenat assays (COVID-19, Mycobacterium tuberculosis - MTB, malaria, leptospirosis), focusing on analytical performance, repeat rates, and time-to-result. Methods:Remnant clinical specimens and commercial controls were analyzed under routine conditions. Performance metrics included accuracy, reproducibility, limit of detection (LOD), linearity, and specificity, when applicable. Each analyte was tested in replicates, and results were compared with established reference methods. Results:The Truenat COVID-19 assay achieved 100% agreement with the reference and LOD of 500 copies/mL. Truenat MTB showed 95% concordance with the comparator, though repeat runs were occasionally required (7.3%). Malaria assays demonstrated high reproducibility and linearity (R2 = 0.9996) with an observed LOD of 3000 copies/mL. Leptospira assays yielded 100% accuracy with a LOD of 750 copies/mL. Performance generally matched manufacturer specifications, though low-burden MTB samples were less consistently detected. Conclusion:Truenat assays showed reliable analytical performance, especially for COVID-19. While MTB detection remains more robust with Xpert Ultra in low-burden cases, Truenat provides a viable point-of-care alternative in resource-limited settings. For malaria and leptospirosis, broader clinical validation is needed before routine implementation in our service.

We report a rare case of polymicrobial late-onset knee prosthetic joint infection (PJI) caused by Parvimonas micra and Staphylococcus aureus. An 80-year-old woman with multiple comorbidities presented, five years after total knee arthroplasty, with progressive pain and radiographic signs of prosthetic loosening. Synovial fluid analysis revealed marked neutrophilic inflammation, and intraoperative tissue cultures, including sonication fluid yielded both pathogens. Identification was confirmed by MALDI-TOF MS and whole-genome sequencing (WGS). The P. micra strain showed 97.2% identity to reference strain SAMN29629855, and carried virulence genes such as groEL, tufA, clpP, ctrD, srtC4, and gaIE, associated with oxidative stress response, adhesion, immune evasion, and biofilm formation. Resistance genes vanW, vanT, and vanY from the van operon were also detected, though vanA and vanB were absent. The patient underwent a two-stage revision surgery and a 12-week course of pathogen-targeted antimicrobial therapy, with complete resolution of symptoms and no recurrence after 12 months. This case highlights the overlooked pathogenicity of P. micra in chronic PJIs, especially in polymicrobial biofilm-related infections. The integration of WGS provided valuable insights into possible genetic characteristics of virulence determinants of this rare cause of PJI.

Introduction: Acute diarrhea is the third leading cause of mortality worldwide in children under 5 years, with approximately 1.7 billion cases per year, according to the World Health Organization. Norovirus stands out as one of the main causes, responsible for 18–20% of all diarrhea cases, with an estimated 685 million cases and more than 200,000 deaths annually. Objective: To describe the main etiological agents of acute diarrhea identified by molecular panel and to characterize the norovirus outbreak in the first quarter of 2025. Methods: Observational, retrospective study from January 1, 2022, to April 30, 2025, using the FilmArray GI®/bioMérieux panel, which detects 22 gastrointestinal pathogens in a single stool sample, performed in a tertiary hospital in São Paulo. Results: Between January 1, 2022, and April 30, 2025, 22,923 tests were performed, with pathogen detection in 36% of samples. The five most frequent agents were: enteropathogenic Escherichia coli (17%), Clostridioides difficile (12%), norovirus GI/GII (11%), enteroaggregative E. coli (9%), and rotavirus (6%). During the norovirus outbreak in the first quarter of 2025, positivity increased to 21.53%, with 48% of cases in the 20–59-year age group and no sex difference (males 51%, females 49%). Conclusion: Epidemiological studies provide crucial information on etiologic agents of acute diarrhea using molecular biology tools, which are important for outbreak surveillance and for monitoring pathogen–age group associations to guide optimal therapeutic management.

Introduction/Objective: Cytomegalovirus (CMV), a member of the Herpesviridae family, is widely distributed globally and usually causes asymptomatic infection or self-limited symptoms, followed by latency for an indeterminate time in immunocompetent individuals. However, in immunosuppressed patients, it can trigger severe clinical manifestations due to reactivation. The emergence of strains resistant to the antivirals ganciclovir, foscarnet, cidofovir, maribavir, and letermovir represents a growing challenge in treatment. In this context, early detection of mutations associated with viral resistance is essential to guide interventions and optimize therapeutic management. This study aimed to develop a molecular test based on next-generation sequencing (NGS) to identify mutations associated with resistance to the main antivirals in key CMV genes, using clinical samples. Methods: Specific primers were designed for the target regions of the UL27, UL51, UL54, UL56, and UL97 genes, where mutations previously associated with antiviral resistance occur. Viral DNA was extracted from clinical samples previously positive for CMV and then submitted to PCR, library preparation with the Microbial Amplicon Prep kit, and sequencing on an Illumina platform. Bioinformatic analysis was conducted with an in-house pipeline, and variants were compared with the HerpesDRG DB v03112024 database. Results: The assay demonstrated 98% sensitivity for detecting resistance variants in samples with viral load ≥1,500 IU/mL and 80% in samples with viral load of 1,000 IU/mL. A total of 39 clinical samples were analyzed, of which 5 showed clinically relevant mutations in the UL97 gene. Mutations in the UL54 and UL56 genes were validated using synthetic DNA. The main mutations identified included C603W, M460V, L595S, and A594V (UL97), I726T, T700A, and Q578H (UL54), and C325F and T244K (UL56), associated with resistance to different antivirals. The assay performance was validated in an international reference laboratory, with advantages such as broader genomic coverage, faster turnaround time, and simultaneous analysis of multiple genes. Conclusion: The developed test is robust, sensitive, and applicable to routine clinical practice for detecting CMV antiviral resistance. Its implementation contributes to personalized management of immunosuppressed patients, optimizing therapeutic choice and reducing the risk of treatment failure.

Background and ObjectivesDengue virus (DENV) infection can cause acute encephalitis. Chronic encephalitis with progressive dementia is rarely reported.MethodsWe present a case of chronic encephalitis with rapidly progressive dementia, in which a potential DENV brain infection was identified with brain tissue metagenomic next-generation sequencing. Brain pathology and molecular diagnosis are also presented.ResultsA 20-year-old man from SP, Brazil, presented with rapidly progressive dementia, speech apraxia, and apathy in June 2022. By January 2023, cognitive testing showed severe global impairment (MMSE score of 18/30). MRI revealed white matter abnormalities and atrophy; CSF analysis disclosed a mild lymphocytic pleocytosis, mildly elevated protein levels, and positive CSF oligoclonal bands. Despite extensive testing ruling out common infectious and inflammatory causes, the patient's condition worsened with executive dysfunction, language impairment, tremors, and myoclonus. In August 2023, a brain biopsy and next-generation sequencing identified DENV-1 genotype V, linked to Brazilian sequences from 2000 to 2022.DiscussionThis case highlights a challenging instance of encephalitis with unknown etiology, where metagenomic analysis detected DENV-1 RNA in brain tissue, suggesting a possible cause.

PURPOSE:Cladophialophora bantiana is a rare cause of severe central nervous system phaeohyphomycosis. RESULTS:A 44-year-old woman from Northeastern Brazil with a history of scotomas, mental confusion, and impaired coordination presented Magnetic Resonance Imaging with vasogenic edema and nodular enhancement in the left occipital lobe. Brain biopsy showed neutrophilic exudate with eosinophils, macrophages, giant multinucleated cells, and septate hyphae with a brown-colored birefringent wall and acute angle branching. Metagenomic analysis of the biopsy's total RNA revealed the presence of RNA sequences highly similari to C. bantiana. Culture confirmed the presence of olivaceous-gray suede-like to floccose colonies, with septate dematiaceous hyphae and long conidia chains from undifferentiated conidiophores, consistent with C. bantiana. CONCLUSIONS:This case highlights the potential of metagenomic testing as a tool for early diagnosis of infections caused by uncommon fungal pathogens. To our knowledge, this is the first report of C. bantiana central nervous system infection in Brazil.

Background:There is a significant concern of the pandemic impact of SARS-CoV-2 infection in immunocompromised patients. These patients can develop long COVID-19 due to impairment of cellular and humoral immunity. On the other hand, prolonged infection can lead to mutations in the SARS CoV-2 genome that can impact on the resistance to antiviral therapy. Remdesivir cases have been reported in patients receiving antiviral drug treatment. Case presentation:A 46-year-old male with previous mantle cell lymphoma resolved by autologous bone marrow transplantation without other comorbidities had SARS-CoV-2 detected in February 2022 and received the recommended antiviral treatment with Remdesivir. COVID-19 evolved in four months with worsening of the symptoms, despite an initial rapid improvements and high RT-PCR Ct values. The patient was relieved from hospital care stable and well but still maintaining positive test results. Conclusions:the patient presented prolonged COVID-19 with persistence of virus detected by RT-PCR for several months. The strain sequenced presented a mutation different from all reported previously. Although it was no possible to sequence the initial strain without these mutations, our data suggests that immunocompromised patient with prolonged COVID-19 may serve as reservoir for strains of SARS-CoV-2 with resistant components in his genome.

Introdução: O vírus da dengue (DENV) é um vírus RNA de sentido positivo pertencente ao gênero Orthoflavivirus, transmitido principalmente pelos mosquitos do gênero Aedes. Existem quatro sorotipos de DENV (DENV1, DENV2, DENV3 e DENV4), cada um com antigenicidade e filogenia distintas. Todos os quatro sorotipos podem causar uma doença com sintomas semelhantes, assim como outras arboviroses. Recentemente, tornaram-se disponíveis ensaios comerciais de qPCR para a detecção do DENV, bem como métodos multiplexados que permitem a detecção simultânea com outros arbovírus como Chikungunya e Zika. Esses métodos apresentam excelente sensibilidade e especificidade diagnóstica, melhorando significativamente a capacidade de diagnóstico. No Brasil, até o mês de maio, foram registrados 4.603.825 casos prováveis de dengue, com 2.451 desses casos evoluindo para óbito. Este número alarmante ressalta a importância de um monitoramento eficaz e contínuo. O monitoramento dos genótipos dos vírus circulantes por sequenciamento é importante por vários motivos, como: variantes patogênicas, levantamento epidemiológico da doença e surgimento de novas linhagens. Objetivo: O presente estudo analisou por Metagenômica de RNA o genoma do vírus da Dengue de cinco pacientes, dos quais quatro estavam internados no nosso serviço. Método: Os pacientes foram diagnosticados através de qPCR e sorologia. Após a extração de RNA, essas amostras foram submetidas a amplificação randômica, preparo de bibliotecas e sequenciamento de nova geração (NGS). Para as análises de Bioinformática um pipeline próprio foi aplicado para categorizar os sorotipos e genótipos. Resultados: Dos cincos pacientes sequenciados, quatro foram identificados como DENV1 - Genótipo V com uma média de cobertura horizontal de 86,2%; e o quinto caso identificado com DENV2 – Genótipo II (Cosmopolitan) apresentando cobertura horizontal de 67%. Conclusão: Os nossos dados são corroborados com base nas análises durante o surto, sobre a predominância dos sorotipos de DENV1 e DENV2. O vírus DENV2 – Genótipo II (Cosmopolitan) é um genótipo emergente, sendo sequenciado pela primeira vez no ano de 2022. Com relação a baixa cobertura horizontal apresentada pelo sorotipo DENV2 – Genótipo II (Cosmopolitan), pode ser explicada pela baixa carga viral. O monitoramento epidemiológico dos sorotipos de Dengue se faz necessário para acompanhamento da doença, bem como avaliar possíveis novos surtos por outros sorotipos.

Introdução: A síndrome hemofagocítica (SH) é uma condição rara caracterizada por uma hiperativação desregulada do sistema imunológico, resultando em hemofagocitose anormal. A SH pode ser dividida em primária e secundária e esta última pode ser desencadeada por patógenos infecciosos, neoplasias, doenças autoimunes, imunossupressão e medicamentos. Objetivo: Entre os gatilhos infecciosos, a família Herpesviridae é responsável por 62% dos casos, com destaque para o Epstein-Barr (43%). Outros herpesvírus, incluindo os tipos 1 e 2, são menos comuns, mais frequente em pacientes imunodeprimidos e raramente afetando adultos imunocompetentes. Método: Relatamos o caso de uma paciente imunocompetente de 27 anos com quadro de SH desencadeada por infecção por vírus herpes simples tipo 1 em nosso serviço. Resultados: A paciente apresentou febre, mialgia, náuseas e vômitos com dois dias de evolução, evoluindo para insuficiência respiratória aguda. A paciente não possuía antecedentes relevantes, imunossupressão ou diagnósticos prévios. Durante a investigação, foram descartadas alterações genéticas associadas à SH primária; tomografia de tórax evidenciava consolidação pulmonar e atelectasias. As sorologias para dengue, Zika, chikungunya, citomegalovírus, Epstein-Barr, clamídia e neisseria foram não reagentes. Apresentou sorologia IgG positiva para herpes simples. Foi então solicitado o PCR em tempo real em lavado broncoalveolar e sangue periférico que confirmou a presença do DNA do herpes simples tipo 1. A paciente continuava apresentando febre além de esplenomegalia, pancitopenia, hiperferritinemia, hipertrigliceridemia e evidência de SH em aspirado de medula óssea. O HScore da paciente indicou uma probabilidade de 98% para SH. O tratamento foi feito com imunoglobulina, corticoterapia e aciclovir evoluindo com reversão dos sinais de disfunção orgânica. Conclusão: A detecção precoce do agente infeccioso foi crucial para o tratamento eficaz. A SH secundária ao herpes vírus tipo 1 é uma condição rara em adultos imunocompetentes, por isso a importância dos testes moleculares no diagnóstico diferencial da etiologia desta síndrome.

Introdução: A Metagenomica shotgun utilizando sequenciamento de nova geração (NGS) possibilita detectar patógenos raros e negligenciados na prática clínica. A técnica desenvolvida em nosso laboratório, utiliza o RNA mensageiro presente na amostra para detecção e identificação dos microrganismos. Objetivo: O objetivo desse estudo foi avaliar o número total de casos testados, porcentagem de positividade e principais patógenos encontrados durante três anos em laboratório de hospital privado terciário. Método: O RNA total é extraído seguido de digestão do DNA e depleção do rRNA/mtRNA. É então realizada a reação de transcrição reversa em duas etapas com primers randômicos, seguido de amplificação por PCR e preparo de biblioteca. As bibliotecas são sequenciadas usando a plataforma Illumina, e em seguida submetida a análise de bioinformática em pipeline desenvolvido internamente. A interpretação de cada resultado é realizada por time multidisciplinar e quando necessário testes ortogonais confirmatórios são realizados. Resultados: Entre janeiro de 2020 até outubro de 2023 foram testados 2373 casos na rotina clínica. Os materiais mais prevalentes foram amostras de plasma e em seguida amostras de liquor. A taxa de positividade geral foi de 21,66%. Os patógenos associados a doenças negligenciadas foram Brucella, arenavirus, leishmania, hantavirus, taenia sp., dengue, chikungunya, monkeypox, vírus da febre amarela, Cladophialophora e hepatite E. Conclusão: A escassez de métodos diagnósticos para patógenos raros e negligenciados pode levar a subnotificação dessas doenças em diversas partes do mundo. Outro fator é que muitas dessas doenças cursam com quadro clínico semelhante, o que dificulta ainda mais o manejo desses pacientes. O teste de metagenômica demonstrou ser eficiente nesse diagnóstico e as taxas de positividade encontradas em nossa população estão de acordo com outros trabalhos publicados.

Purpose: The microbiology pattern of neonatal conjunctivitis has changed over time, and the incidence of gonococcal conjunctivitis is almost nil. This study aimed to determine the etiology of neonatal conjunctivitis cases referred to a tertiary health center in Brazil. Methods: From 2017 to 2020, conjunctival swabs were taken from neonates with clinical signs of conjunctivitis and tested with bacterial culture and polymerase chain reaction for Neisseria gonorrhoeae and Chlamydia trachomatis. Results: A total of 51 neonates were included in the 3-year study. Chlamydial conjunctivitis was diagnosed in 39 (76.5%) patients, and microbial growth was detected in 13 (25.5%) patients. The most isolated bacterium was Staphylococcus epidermidis (n=6, 11.8%), followed by other coagulase-negative Staphylococcus species (n=4, 7.8%) and S. aureus (n=2, 3.9%). One S. aureus isolate was resistant to oxacillin. There were no cases of gonococcal conjunctivitis. Ten (19.6%) patients showed polymerase chain reaction-negative C. trachomatis and negative bacterial culture test results. Conclusion: Findings show that C. trachomatis is the most common pathogen causing neonatal conjunctivitis. The high prevalence of C. trachomatis infection highlights the importance of screening and treating pregnant woman.

We describe a case of a 33-year-old male presented with fever, myalgia, nausea, and asthenia for six days. The patient lived in a rural area. Initial hypotheses included arbovirus infection, viral hepatitis, and Lyme disease. Reverse transcriptase polymerase chain reaction (RT-PCR) tests for Dengue, Zika, and Chikungunya resulted negative. We were able to recover complete S, L, and M segments of virus in the Orthohantavirus genome.

BACKGROUND CONTEXT:Reports of Cutibacterium acnes isolated in cultures of intervertebral disc samples suggest it as possibly responsible for inflammatory conditions causing Modic changes on spinal magnetic resonance imaging (MRI). PURPOSE:Our objective was to investigate the prevalence of C. acnes in samples of intervertebral disc of patients with lumbar disc herniation; to investigate prognostic factors and the relationship of Modic changes with infection 1 year after microdiscectomy. STUDY DESIGN:Prospective cohort study. PATIENT SAMPLE:In this single-center study, patients consecutively operated on for disc herniation had samples of the disc, multifidus muscle and ligamentum flavum (as an indication of contamination) extracted for culture. OUTCOME MEASURES:Age, sex, alcohol and tobacco consumption, body mass index; function, pain, and Modic chances in MRI before surgery and MRI 1 year later; rate of disc, muscle and ligament infection (primary outcome); diabetes and corticoid use (confoundings). METHODS:The protruded disc, muscle and ligament samples were sent for culture analysis in up to 30 minutes. A subsample of 17 patients underwent next-generation sequencing (NGS) molecular analysis too. We performed descriptive analysis and comparison of groups of patients with and without infection or contamination using Student's t, Mann-Whitney, chi-square, or Fisher's exact tests as appropriate, and pre- and postsurgical comparisons with the Wilcoxon test. RESULTS:From January 2018 to September 2019, 112 patients underwent open lumbar microdiscectomy, 67 (59.8%) men. Cultures showed 7 (6.3%) positive cases in the disc (2 with C. acnes), 3 (2.7%) in the ligament, and 12 (10, 7%) in muscle. No evidence of a difference in Modic alterations pre- or postoperatively was found between patients with and without positive culture 1 year after surgery. No association was found between culture positivity and functional or pain differences either. NGS results were all negative for C. acnes. CONCLUSIONS:We identified infective bacterial presence in the herniated disc in less than 2% of patients with disc herniation. C. acnes was not identified in any disc microbiome analysis. No significant association was observed between positivity for tissue infection and any clinical prognostic factor.

A 68-year-old Brazilian woman had 3 months of progressive fatigue, difficulty walking and 18 kg weight loss. On examination, there was gait apraxia and executive dysfunction. MR scan of brain showed communicating hydrocephalus and a cerebrospinal fluid showed 105 white cells/& mu;L (& LE;5), predominantly lymphocytes, protein of 1.35 g/L (0.15-0.45) and the glucose content of 0.06 mmol/L (3.3-4.4). We suspected an infective cause and used of metagenomic next-generation sequencing to diagnose neurocysticercosis. This case highlights the challenge of diagnosing chronic meningitis and the relevance of genetic approaches in diagnosing neurological infections.

Background: In the initial phases of the COVID-19 pandemic, strategies adopted to reduce the dissemination of SARS-CoV-2 relied on non-pharmacological interventions, including physical distancing. Mobility restrictions affected the availability and quality of care for many health conditions, including sexually transmitted infections. Objective: To investigate the impact of the COVID-19 pandemic mobility restriction on syphilis and HIV testing in outpatient settings. Methods: In this study, we collected the weekly number of syphilis and HIV tests performed in a referent laboratory in Sao Paulo, Brazil, as well as the percentage of positive tests between January 2019 and December 2021. We also retrieved data on retail and recreation mobility in Sao Paulo city using Google COVID-19 Community Mobility Reports. We explored the associa-tion between populational mobility and the number of weekly tests and the association between the number of weekly tests and the percentage of positive results during the pandemic period. The analyses were conducted separately for syphilis and HIV tests. Results: We found that mobility restrictions during the COVID-19 pandemic have been asso-ciated with a significant decrease in both syphilis and HIV tests performed in outpatient settings. We also observed that the number of tests performed was inversely associated with the percentage of positive results for syphilis; this association was also apparent for HIV tests in the first wave of the pandemic in the graphic analysis. Conclusion: Taken together, our findings suggest an indirect impact of COVID-19 pandemic -related mobility restrictions on the uptake of diagnostic tests for HIV and syphilis and the potential adoption of targeted-testing strategies. Understanding the extent and complexity of COVID-19 aftermaths on specific conditions and communities is essential to build strate-gies to mitigate the long-term consequences of COVID-19. (c) 2023 Sociedade Brasileira de Infectologia. Published by Elsevier Espana, S.L.U. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Cycle threshold (Ct) values in COVID-19 reverse-transcription polymerase chain reaction (RT-PCR) tests estimate the viral load in biological samples. Studies have investigated variables associated with SARS-CoV-2 viral load, aiming to identify factors associated with higher transmissibility. Using the results from tests performed between May/2020-July/2022 obtained from the database of a referent hospital in Sao Paulo, Brazil, we investigated associations between Ct values and patient's age, gender, sample collection setting and pandemic period according to the predominant SARS-CoV-2 variant locally. We also examined variations in Ct values, COVID-19 incidence, mortality, and vaccination coverage over time. The study sample included 42,741 tests. Gender was not significantly associated with Ct values. Age, sample collection setting and the pandemic period were significantly associated with Ct values even after adjustment to the multivariable model. Results showed lower Ct values in older groups, during the Gamma and Delta periods, and in samples collected in emergency units; and higher Ct values in children under 10 years old, home-based tests, during the Omicron period. We found evidence of a linear trend in the association between age and Ct values, with Ct values decreasing as age increases. We found no clear temporal associations between Ct values and local indicators of COVID-19 incidence, mortality, or vaccination between February/2020-November/2022. Our findings suggest that SARS-CoV-2 Ct values, a proxy for viral load and transmissibility, can be influenced by demographic and epidemiological variables.

BACKGROUND CONTEXT There is apparent causality between chronic infection of the intervertebral disc and its degenerative process. Although disc is considered a sterile tissue, collected samples of uninfected patients sent to culture testing resulted positive. PURPOSE The purpose of this study was to analyze the microbiome of the intervertebral disc by using and validating the next-generation sequencing (NGS) molecular test, controlled with tissue culture and clinical presentation of patients. STUDY DESIGN/SETTING Prospective study of consecutive patients in a hospital. PATIENT SAMPLE Patients with lumbar disc herniation undergoing open microdiscectomy aging 18 to 65 years. OUTCOME MEASURES NGS, tissue culture METHODS Subjects undergoing open decompression surgery for lumbar disc herniation were consecutively included and clinically followed for one year. Three samples of the excised herniated disc fragment were sent to tissue culture and another sample of the disc was sent to NGS test for microbiome analysis. Control samples of the ligamentum flavum and deep muscle were collected and sent to culture. RESULTS A total of 17 patients were included. All patients presented negative cultures of the removed disc samples, as well as negative cultures of muscle and yellow ligament. None of the patients evolved to clinical infection one year after surgery, nor presented significant alteration of laboratory markers. NGS mapped a mean of 14,645 (range 6,540 to 27,176) DNA sequences for each disc sample of each patient. There were a total of 45 different bacteria genera remnants with different amount of DNA sequences detected. There was a mean of 8 (range 3-17) different bacterial elements in each sample of intervertebral disc. Three bacteria were present in all disc samples (Herbaspirillum, Ralstonia, and Burkolderia). Although there were a considerable mean number of bacterial sequences mapped in each disc sample, the amount of sequences related to bacteria was low. Cutibacterium acnes elements was not found in any disc microbiome analysis. CONCLUSIONS NGS has been proven to adequately determine bacterial DNA presence within the intervertebral disc. C. acnes was not isolated in culture neither in microbiome analysis of patients with lumbar disc herniation. We cannot confirm disc sterility since, even if it does not cause infection, there is bacterial or remnant DNA in herniated discs.

COVID‐19 patients on mechanical ventilation are at risk to develop invasive aspergillosis. To provide additional data regarding this intriguing entity, we conducted a retrospective study describing risk factors, radiology and prognosis of this emerging entity in a Brazilian referral centre.

Introdução: A identificação laboratorial oportuna é uma etapa essencial para o diagnóstico e tratamento precoces, e para interromper cadeias de transmissão de infecções sexualmente transmissíveis (IST). Alterações no padrão de mobilidade humana e realocações de recursos diagnósticos e terapêuticos em decorrência pandemia da COVID-19 vêm afetando diversos aspectos do cuidado a outras doenças, tais como as sífilis. A redução de testagens pode associar-se a testagem direcionada a indivíduos com maior risco de infecção, resultando em aumento da porcentagem de exames positivos. Objetivo: Descrever o efeito da pandemia da COVID-19 sobre o número de testes solicitados para sífilis, e sobre a porcentagem de testes positivos. Correlacionar o número de testes solicitados e porcentagem de positivos em cada quinzena no período de janeiro de 2019 a outubro de 2021 com mensurações da mobilidade populacional utilizando a ferramenta Google Mobility. Método: Nesse estudo de série temporal, extraímos do banco de dados do laboratório do Hospital Albert Einstein dados sobre a frequência quinzenal de testes diagnósticos para sífilis (quimiluminescência), bem como seus resultados, descrevendo as porcentagens de testes positivos, no período de janeiro de 2019 a outubro de 2021. Análise foi realizada por métodos gráficos e testes de correlação de Spearman. Resultados: 41773 testes de quimiluminescência foram incluídos na análise. A avaliação gráfica revelou uma queda acentuada no número de testes solicitados no período inicial da pandemia (março-maio de 2020), e um aumento moderado entre maio e setembro de 2021, coincidentes respectivamente com a redução e retomada da mobilidade populacional. Conforme esperado, a redução da testagem acompanhou-se de aumento da porcentagem de exames positivos, com uma positividade média de 8,3% no período pré-pandemia para 13,4% no período de março-maio 2020. Observamos correlação estatisticamente significante entre testagem e mobilidade (Rho = 0,594, p < 0,0001), e correlação inversa entre testagem e porcentagem de exames positivos (Rho = -0,517, p = 0,0005). Conclusão: Alterações da mobilidade populacional e alocação de recursos decorrentes da pandemia da COVID-19 ou outros eventos excepcionais podem ter impacto negativo sobre o cuidado a outras doenças, tais como a sífilis. Estratégias para ampliar a capacidade de testagem, incluindo o uso de autotestes, podem favorecer o acesso ao diagnóstico e tratamento da sífilis e outras IST.