Assessment of the Accuracy, Usability and Acceptability of a Rapid Test for the Simultaneous Diagnosis of Syphilis and HIV Infection in a Real-Life Scenario in the Amazon Region, Brazil.

We field-assessed the accuracy, acceptability, and feasibility of the SD BIOLINE HIV/Syphilis Duo rapid diagnostic test in three groups: pregnant women, female sex workers (FSW), and men who have sex with men (MSM). Venous blood samples collected in the field were compared with the respective gold standard methods: SD BIOLINE HIV/Syphilis Duo Treponemal Test versus FTA-abs (Wama brand) treponemal laboratory test for syphilis, and SD BIOLINE HIV/Syphilis Duo Test versus the fourth generation Genscreen Ultra HIV Ag-Ag (Bio-Rad brand) laboratory test for HIV. From a total of 529 participants, 397 (75.1%) were pregnant women, 76 (14.3%) FSW and 56 (10.6%) MSM. Sensitivity and specificity parameters of HIV were 100.0% (95% CI: 82.35-100.0%) and 100.0% (95% CI: 99.28-100.0%), respectively. Sensitivity and specificity parameters found for TP antibody detection were 95.00% (95% CI: 87.69-98.62%) and 100.0% (95% CI: 98.18-100.0%), respectively. The SD BIOLINE HIV/Syphilis Duo Test showed high acceptability among participants (85.87%) and health professionals (85.51%), as well as easy usability by professionals (91.06%). The usability of the SD BIOLINE HIV/Syphilis Duo Test kit would not be a barrier to accessing rapid testing, if the product were incorporated into the list of health service supplies.

GyrB in silico mining in 27 151 global gonococcal genomes from 1928-2021 combined with zoliflodacin in vitro testing of 71 international gonococcal isolates with different GyrB, ParC and ParE substitutions confirms high susceptibility.

OBJECTIVES: Antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a global threat and novel treatment alternatives are imperative. Herein, susceptibility to the novel antimicrobial zoliflodacin, currently in a global Phase 3 randomized controlled clinical trial for gonorrhoea treatment, was investigated by screening for zoliflodacin GyrB target mutations in publicly available gonococcal genomes and, where feasible, determination of the associated zoliflodacin MIC. METHODS: The European Nucleotide Archive was queried using the search term 'Taxon: 485'. DNA sequences from 27 151 gonococcal isolates were analysed and gyrB, gyrA, parC and parE alleles characterized. RESULTS: GyrB amino acid alterations were rare (97.0% of isolates had a wild-type GyrB sequence). GyrB V470L (2.7% of isolates) was the most prevalent alteration, followed by S467N (0.12%), N. meningitidis GyrB (0.092%), V470I (0.059%), Q468R/P (0.015%), A466T (0.0074%), L425I + L465I (0.0037%), L465I (0.0037%), G482S (0.0037%) and D429V (0.0037%). Only one isolate (0.0037%) carried a substitution in a resistance-associated GyrB codon (D429V), resulting in a zoliflodacin MIC of 8 mg/L. None of the other detected gyrB, gyrA, parC or parE mutations caused a zoliflodacin MIC outside the wild-type MIC distribution. CONCLUSIONS: The zoliflodacin target GyrB was highly conserved among 27 151 global gonococcal isolates cultured in 1928-2021. The single zoliflodacin-resistant clinical isolate (0.0037%) was cultured from a male patient in Japan in 2000. Evidently, this strain has not clonally expanded nor has the gyrB zoliflodacin-resistance mutation disseminated through horizontal gene transfer to other strains. Phenotypic and genomic surveillance, including gyrB mutations, of zoliflodacin susceptibility are imperative.

In vitro selection of Neisseria gonorrhoeae unveils novel mutations associated with extended-spectrum cephalosporin resistance.

The emergence of Neisseria gonorrhoeae strains resistant to extended-spectrum cephalosporins (ESCs) is a worldwide concern because this class of antibiotics represents the last empirical treatment option for gonorrhea. The abusive use of antimicrobials may be an essential factor for the emergence of ESC resistance in N. gonorrhoeae. Cephalosporin resistance mechanisms have not been fully clarified. In this study, we mapped mutations in the genome of N. gonorrhoeae isolates after resistance induction with cefixime and explored related metabolic pathways. Six clinical isolates with different antimicrobial susceptibility profiles and genotypes and two gonococcal reference strains (WHO F and WHO Y) were induced with increasing concentrations of cefixime. Antimicrobial susceptibility testing was performed against six antimicrobial agents before and after induction. Clinical isolates were whole-genome sequenced before and after induction, whereas reference strains were sequenced after induction only. Cefixime resistance induction was completed after 138 subcultures. Several metabolic pathways were affected by resistance induction. Five isolates showed SNPs in PBP2. The isolates M111 and M128 (ST1407 with mosaic penA-34.001) acquired one and four novel missense mutations in PBP2, respectively. These isolates exhibited the highest minimum inhibitory concentration (MIC) for cefixime among all clinical isolates. Mutations in genes contributing to ESC resistance and in other genes were also observed. Interestingly, M107 and M110 (ST338) showed no mutations in key determinants of ESC resistance despite having a 127-fold increase in the MIC of cefixime. These findings point to the existence of different mechanisms of acquisition of ESC resistance induced by cefixime exposure. Furthermore, the results reinforce the importance of the gonococcal antimicrobial resistance surveillance program in Brazil, given the changes in treatment protocols made in 2017 and the nationwide prevalence of sequence types that can develop resistance to ESC.

Genetic diversity and antimicrobial resistance of invasive, noninvasive and colonizing group B Streptococcus isolates in southern Brazil.

Introduction: Group B Streptococcus (GBS) is a human commensal bacterium that is also associated with infection in pregnant and non-pregnant adults, neonates and elderly people. Gap Statement: The authors hypothesize that knowledge of regional GBS genetic patterns may allow the use of prevention and treatment measures to reduce the burden of streptococcal disease. Aim: The aim was to report the genotypic diversity and antimicrobial sensitivity profiles of invasive, noninvasive urinary and colonizing GBS strains, and evaluate the relationships between these findings. Methodology: The study included consecutive and non-duplicated GBS isolates recovered in southern Brazil from 2015 to 2017. We performed multiple-locus variable-number tandem repeat analysis (MLVA) and PCR analyses to determine capsular serotypes and identify the presence of the resistance genes mefA/E, ermB and ermA/TR, and also antibiotic susceptibility testing. Results: The sample consisted of 348 GBS strains, 42 MLVA types were identified, and 4 of them represented 64 % of isolates. Serotype Ia was the most prevalent (42.2 %) and was found in a higher percentage associated with colonization, followed by serotypes V (24.4 %), II (17.8 %) and III (7.8 %). Serotype V was associated with invasive isolates and serotypes II and III with noninvasive isolates, without significant differences. All isolates were susceptible to penicillin. GBS 2018/ hvgA was observed in 17 isolates, with 11 belonging to serogroup III. The Hunter-Gaston diversity index was calculated as 0.879. The genes mefA/E, erm/B and erm/A/TR were found in 45, 19 and 46 isolates. Conclusion: This report suggests that the circulating GBS belong to a limited number of genetic lineages. The most common genotypes were Ia/MT12 and V/MT18, which are associated with high resistance to macrolides and the presence of the genes mefA/E and ermA/TR. Penicillin remains the antibiotic of choice. Implementation of continuous surveillance of GBS infections will be essential to assess GBS epidemiology and develop accurate GBS prevention, especially strategies associated with vaccination.

National surveillance of Neisseria gonorrhoeae antimicrobial susceptibility and epidemiological data of gonorrhoea patients across Brazil, 2018-20.

Objectives: To (i) describe the nationwide antimicrobial susceptibility of Neisseria gonorrhoeae (NG) isolates cultured across Brazil in 2018-20 and compare it with NG antimicrobial resistance data from 2015-16, and (ii) present epidemiological data of the corresponding gonorrhoea patients in 2018-20. Methods: Twelve representative sentinel sites cultured NG isolates from men with urethral discharge. Susceptibility to eight antimicrobials was examined using agar dilution method, according to WHO standards. The consenting participants were invited to provide epidemiological data. Results: In total, 633 NG isolates (one isolate per participant) were analysed, and 449 (70.9%) questionnaires were answered. Heterosexual (68.2%) and homosexual (23.1%) sexual orientations were common, and most prevalent types of unprotected sexual intercourse were vaginal insertive (69.9%), oral giving (56.6%) and anal insertive (47.4%). The levels of in vitro NG resistance to ciprofloxacin, tetracycline, benzylpenicillin, azithromycin, cefixime, gentamicin, spectinomycin and ceftriaxone were 67.3%, 40.0%, 25.7%, 10.6%, 0.3%, 0%, 0% and 0%, respectively. Compliance with the recommended first-line ceftriaxone 500 mg plus azithromycin 1 g therapy was high (90.9%). Conclusions: Compared with 2015-16, ciprofloxacin resistance has remained high and azithromycin and cefixime resistance rates have increased in Brazil. Resistance remained lacking to ceftriaxone, gentamicin and spectinomycin, which all are gonorrhoea treatment options. The increasing azithromycin resistance in Brazil and internationally may threaten the future use of azithromycin in dual regimens for treatment of gonorrhoea. Consequently, continued and enhanced quality-assured surveillance of gonococcal AMR, and ideally also treatment failures and including WGS, is imperative in Brazil and worldwide.

Spectinomycin, gentamicin, and routine disc diffusion testing: An alternative for the treatment and monitoring of multidrug-resistant Neisseria gonorrhoeae?

INTRODUCTION: Neisseria gonorrhoeae is a major concern of public health due to its extraordinary capacity to develop and acquire resistance to different antimicrobials used to treat gonorrhoea. Limited treatment options and uncontrolled transmission have raised the need to assess the antimicrobial susceptibility profile of the isolates and to establish affordable alternatives for laboratory diagnosis. OBJECTIVES: This study aimed to (i) determine the susceptibility profile of 336 clinical isolates of N. gonorrhoeae to ceftriaxone, azithromycin, ciprofloxacin, spectinomycin and gentamicin by the gold standard agar dilution method; (ii) assess the agreement among agar dilution and disc diffusion results for ciprofloxacin, azithromycin, ceftriaxone, spectinomycin and gentamicin. RESULTS: All isolates were susceptible to ceftriaxone and spectinomycin. The levels of resistance to azithromycin and ciprofloxacin were 3.9% and 35.1%, respectively. Intermediate susceptibility to gentamicin was observed in 19.4% of isolates. There was 100% agreement between methods for spectinomycin and ceftriaxone, 99.7% for ciprofloxacin, and 85.7% for azithromycin. For gentamicin, there was 86.3% agreement between agar dilution and disc diffusion, resulting in intermediate susceptible by one method and susceptible by the other method, defined as minor errors. The discordance among agar dilution and disc diffusion results is acceptable for ciprofloxacin, ceftriaxone and spectinomycin as per CLSI M23-Ed4. CONCLUSIONS: Spectinomycin and gentamicin can be considered in some cases as options for the treatment of gonorrhoea in Brazil. Disc diffusion can be an alternative method in routine testing with comparable accuracy to agar dilution.

Emergence of Two Distinct SARS-CoV-2 Gamma Variants and the Rapid Spread of P.1-like-II SARS-CoV-2 during the Second Wave of COVID-19 in Santa Catarina, Southern Brazil.

The western mesoregion of the state of Santa Catarina (SC), Southern Brazil, was heavily affected as a whole by the COVID-19 pandemic in early 2021. This study aimed to evaluate the dynamics of the SARS-CoV-2 virus spreading patterns in the SC state from March 2020 to April 2021 using genomic surveillance. During this period, there were 23 distinct variants, including Beta and Gamma, among which the Gamma and related lineages were predominant in the second pandemic wave within SC. A regionalization of P.1-like-II in the Western SC region was observed, concomitant to the increase in cases, mortality, and the case fatality rate (CFR) index. This is the first evidence of the regionalization of the SARS-CoV-2 transmission in SC and it highlights the importance of tracking the variants, dispersion, and impact of SARS-CoV-2 on the public health systems.

Genomic analysis of Neisseria elongata isolate from a patient with infective endocarditis.

Neisseria elongata is part of the commensal microbiota of the oropharynx. Although it is not considered pathogenic to humans, N. elongata has been implicated in several cases of infective endocarditis (IE). Here, we report a case of IE caused by N. elongata subsp. nitroreducens (Nel_M001) and compare its genome with 17 N. elongata genomes available in GenBank. We also evaluated resistance and virulence profiles with Comprehensive Antibiotic Resistance and Virulence Finder databases. The results showed a wide diversity among N. elongata isolates. Based on the pangenome cumulative curve, we demonstrate that N. elongata has an open pangenome. We found several different resistance genes, mainly associated with antibiotic efflux pumps. A wide range of virulence genes was observed, predominantly pilus formation genes. Nel_M001 was the only isolate to present two copies of some pilus genes and not present nspA gene. Together, our results provide insights into how this commensal microorganism can cause IE and may assist further biological investigations on nonpathogenic Neisseria spp. Case reporting and pangenome analyses are critical for enhancing our understanding of IE pathogenesis, as well as for alerting physicians and microbiologists to enable rapid identification and treatment to avoid unfavorable outcomes.

An investigation about chronic prostatitis in ankylosing spondylitis.

BACKGROUND: Chronic prostatitis has been a common disease reported with high frequency in ankylosing spondylitis (AS) even from decades ago. Infectious (Chlamydia trachomatis) or non-infectious (uric acid) prostatitis can hypothetically trigger vertebral inflammation in AS. This study aimed to assess the features of chronic prostatitis in patients with AS compared to healthy controls. METHODS: A cross-sectional study including male patients with AS and healthy controls who agreed to undergo a prostate examination was conducted. Structured clinical interviews, prostate physical examinations, and cytological, biochemical, and microbiological tests on urinary samples collected before and after standardized prostatic massage (pre- and post-massage test) were performed. RESULTS: Ninety participants (45 AS patients, mean age: 52.5 ± 10.0 years, with longstanding disease, 12.4 ± 6.9 years, and 45 controls, mean age: 52.8 ± 12.1 years) were included. National Institutes of Health - Chronic Prostatitis Symptom Index (NIH-CPSI) scores were similar in the AS and control groups (4.0 [1.0-12.0] vs. 5.0 [1.0-8.5], p = 0.994). The frequencies of symptoms of chronic prostatitis (NIH-CPSI Pain Domain ≥4) were also similar in both groups (23.3% vs. 22.7%, p = 0.953). Results of polymerase chain reaction tests for Chlamydia trachomatis were negative in all tested urinary samples, and uric acid concentrations and leukocyte counts were similar in all pre- and post-massage urinary samples. CONCLUSIONS: In this study, chronic prostatitis occurred in male patients with AS, but its frequency and characteristics did not differ from those found in the healthy male population of similar age.

Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. This article presents direct tests to detect Treponema pallidum in lesions and algorithms that combine treponemal and non-treponemal antibody tests to assist in syphilis diagnosis, with the aim of contributing to the efforts of health service managers and health professionals in qualifying health care. The article also covers the use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring the treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The critical role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted. Highlighted excerpt: During the natural evolution of syphilis, activity periods with distinct clinical, immunological, and histopathological characteristics are interspersed with latent periods when there are no signs or symptoms, making access to tests critical for early diagnosis.

Brazilian Protocol for Sexually Transmitted Infections 2020: human papillomavirus (HPV) infection.

This article addresses human papillomavirus (HPV) infection, one of the topics covered by the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health. The Protocol and Guidelines have been developed based on scientific evidence and validated in discussions with specialists. This article presents epidemiological and clinical aspects and guidelines for health service managers and health workers about diagnosing and treating people with papillomavirus infection. This theme is a significant public health issue since it is the most prevalent sexually transmitted infection globally, capable of triggering the oncogenic process of cervical cancer and the possibility of anogenital warts. Important information is presented for gaining knowledge about HPV and action strategies for infection prevention and control, provision of quality care, and effective treatment of the disease. Infection by the human papillomavirus is the sexually transmitted infection of the highest contagion, superior to genital herpes and HIV.

[Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests]. / Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

[Brazilian Protocol for Sexually Transmitted Infections 2020: human papillomavirus (HPV) infection]. / Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: infecção pelo papilomavírus humano (HPV).

This article addresses human papillomavirus (HPV) infection, this being one of the topics covered by the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health. The Protocol and Guidelines have been developed based on scientific evidence and validated in discussions with specialists. This article presents epidemiological and clinical aspects, as well as guidelines for health service managers and health workers about diagnosing and treating people with papillomavirus infection. This theme is an important public health problem, since it is the most prevalent sexually transmitted infection in the world, capable of triggering the oncogenic process of cervical cancer, as well as the possibility anogenital warts occurring. Important information is presented for gaining knowledge about HPV, as well as action strategies for infection prevention and control, provision of quality care and effective treatment of the disease.

O artigo aborda a infecção pelo papilomavírus humano (human papillomavirus, HPV), tema constitutivo do Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis, publicado pelo Ministério da Saúde. Tal documento foi elaborado com base em evidências científicas e validado em discussões e consenso entre especialistas. São apresentados aspectos epidemiológicos e clínicos, bem como orientações para os gestores e profissionais de saúde no diagnóstico, tratamento e prevenção da infecção pelo HPV. Este tema representa importante problema de saúde pública, haja vista essa infecção sexualmente transmissível ser a mais prevalente no mundo, capaz de desencadear o processo oncogênico do câncer do colo uterino, além de possibilitar a ocorrência de verrugas anogenitais. Neste artigo, são apresentadas informações importantes para o conhecimento do HPV, estratégias de ação para a prevenção e controle da infecção, uma assistência de qualidade e tratamento efetivo da doença.

El tema del papilomavirus humano (PVH) es uno de los capítulos del Protocolo Clínico y Conductas Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual, publicado por el Ministerio de Salud de Brasil. Este documento fue desarrollado en base a evidencia científica y validado en discusiones con especialistas. Se presentan aspectos epidemiológicos y clínicos, así como guías para gestores y profesionales de la salud en el diagnóstico, tratamiento y prevención de la infección por papilomavirus. Este tema constituye uno de los principales problemas de salud pública, pues además de ser la infección transmitida sexualmente más prevalente en todo el mundo, puede tener implicaciones para el proceso oncogénico del cáncer de cuello uterino y la posibilidad de presentar verrugas anogenitales. Se presenta información sobre estrategias de acciones de prevención y control, las cuales son importantes para entender el problema, ofrecer asistencia de calidad y tratamiento efectivo.

The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019.

Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ± 0.02 log10 SARS-CoV-2 genome copies (GC) L-1), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ± 0.12 log10 SARS-CoV-2 GC L-1, ranging from 5.49 ± 0.02 log10 GC L-1 (27th November 2019) to 6.68 ± 0.02 log10 GC L-1 (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).

Swab pooling: A new method for large-scale RT-qPCR screening of SARS-CoV-2 avoiding sample dilution.

To minimize sample dilution effect on SARS-CoV-2 pool testing, we assessed analytical and diagnostic performance of a new methodology, namely swab pooling. In this method, swabs are pooled at the time of collection, as opposed to pooling of equal volumes from individually collected samples. Paired analysis of pooled and individual samples from 613 patients revealed 94 positive individuals. Having individual testing as reference, no false-positives or false-negatives were observed for swab pooling. In additional 18,922 patients screened with swab pooling (1,344 pools), mean Cq differences between individual and pool samples ranged from 0.1 (Cr.I. -0.98 to 1.17) to 2.09 (Cr.I. 1.24 to 2.94). Overall, 19,535 asymptomatic patients were screened using 4,400 RT-qPCR assays. This corresponds to an increase of 4.4 times in laboratory capacity and a reduction of 77% in required tests. Therefore, swab pooling represents a major alternative for reliable and large-scale screening of SARS-CoV-2 in low prevalence populations.

Serum Complement Activation by C4BP-IgM Fusion Protein Can Restore Susceptibility to Antibiotics in Neisseria gonorrhoeae.

Neisseria gonorrhoeae is the etiological agent of gonorrhea, the second most common bacterial sexually transmitted infection worldwide. Reproductive sequelae of gonorrhea include infertility, ectopic pregnancy and chronic pelvic pain. Most antibiotics currently in clinical use have been rendered ineffective due to the rapid spread of antimicrobial resistance among gonococci. The developmental pipeline of new antibiotics is sparse and novel therapeutic approaches are urgently needed. Previously, we utilized the ability of N. gonorrhoeae to bind the complement inhibitor C4b-binding protein (C4BP) to evade killing by human complement to design a chimeric protein that linked the two N-terminal gonococcal binding domains of C4BP with the Fc domain of IgM. The resulting molecule, C4BP-IgM, enhanced complement-mediated killing of gonococci. Here we show that C4BP-IgM induced membrane perturbation through complement deposition and membrane attack complex pore insertion facilitates the access of antibiotics to their intracellular targets. Consequently, bacteria become more susceptible to killing by antibiotics. Remarkably, C4BP-IgM restored susceptibility to azithromycin of two azithromycin-resistant clinical gonococcal strains because of overexpression of the MtrC-MtrD-MtrE efflux pump. Our data show that complement activation can potentiate activity of antibiotics and suggest a role for C4BP-IgM as an adjuvant for antibiotic treatment of drug-resistant gonorrhea.

Genomic characterization of variants on mycolic acid metabolism genes in Mycobacterium tuberculosis isolates from Santa Catarina, Southern Brazil.

Mycobacterium tuberculosis has a complex cell wall containing mycolic acids (MA), which play an important role in pathogenesis, virulence, and survival by protecting the cell against harsh environments. Studies have shown that genes encoding enzymes involved in MA synthesis are essential to mycobacterial functionality. Here, we used whole-genome sequencing to evaluate mutations in genes related to MA metabolism in M. tuberculosis isolates from pulmonary tuberculosis patients of the Florianópolis Metropolitan Area, Santa Catarina, Brazil, and assessed associations with clinical, epidemiological, and genotypic data. The mutations Rv3057c Asp112Ala (104/151), Rv3720 His70Arg (104/151), and Rv3802c Val50Phe (105/151) were identified in about 69% of the isolates and were related to the LAM lineage. SIT 216/LAM5 (13.2%, 20/151) had the highest frequency and presented the mutations accD2 Lys23Glu, kasA Gly269Ser, mmaA4 Asn165Ser, otsB1 Asp617Asn, Rv3057c Asp112Ala, Rv3720 His70Arg, Rv3802c Val50Phe, and tgs4 Ala216Glu. All SIT 73/T isolates (6.6%, 10/151) showed a characteristic and exclusive gene mutation pattern: amiD Rv3376 3790075G > A, fbpA-aftB 4266941G > A, echA11 Asn220fs, and otsB2 Ser110Arg. SITs 20/LAM1, 64/LAM6, 50/H3, 137/X2, and 119/X1 were also related to specific mutations. SITs from the LAM lineage differed in mutation profile from those of the T, Haarlem, and X lineages. Isolates from patients who had treatment failure showed mutations that do not seem to have a pattern related to this outcome. It was possible to identify a broad repertoire of single-nucleotide polymorphisms in genes related to MA metabolism in M. tuberculosis isolates. This study also described, for the first time, the variability between different SITs/sublineages of Lineage 4 circulating in Florianópolis Metropolitan Area.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: infecção pelo papilomavírus humano (HPV) / Brazilian Protocol for Sexually Transmitted Infections 2020: human papillomavirus (HPV) infection / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: infección por papilomavirus humano (PVH)

Resumo O artigo aborda a infecção pelo papilomavírus humano (human papillomavirus, HPV), tema constitutivo do Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis, publicado pelo Ministério da Saúde. Tal documento foi elaborado com base em evidências científicas e validado em discussões e consenso entre especialistas. São apresentados aspectos epidemiológicos e clínicos, bem como orientações para os gestores e profissionais de saúde no diagnóstico, tratamento e prevenção da infecção pelo HPV. Este tema representa importante problema de saúde pública, haja vista essa infecção sexualmente transmissível ser a mais prevalente no mundo, capaz de desencadear o processo oncogênico do câncer do colo uterino, além de possibilitar a ocorrência de verrugas anogenitais. Neste artigo, são apresentadas informações importantes para o conhecimento do HPV, estratégias de ação para a prevenção e controle da infecção, uma assistência de qualidade e tratamento efetivo da doença.

Abstract This article addresses human papillomavirus (HPV) infection, this being one of the topics covered by the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health. The Protocol and Guidelines have been developed based on scientific evidence and validated in discussions with specialists. This article presents epidemiological and clinical aspects, as well as guidelines for health service managers and health workers about diagnosing and treating people with papillomavirus infection. This theme is an important public health problem, since it is the most prevalent sexually transmitted infection in the world, capable of triggering the oncogenic process of cervical cancer, as well as the possibility anogenital warts occurring. Important information is presented for gaining knowledge about HPV, as well as action strategies for infection prevention and control, provision of quality care and effective treatment of the disease.

Resumen El tema del papilomavirus humano (PVH) es uno de los capítulos del Protocolo Clínico y Conductas Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual, publicado por el Ministerio de Salud de Brasil. Este documento fue desarrollado en base a evidencia científica y validado en discusiones con especialistas. Se presentan aspectos epidemiológicos y clínicos, así como guías para gestores y profesionales de la salud en el diagnóstico, tratamiento y prevención de la infección por papilomavirus. Este tema constituye uno de los principales problemas de salud pública, pues además de ser la infección transmitida sexualmente más prevalente en todo el mundo, puede tener implicaciones para el proceso oncogénico del cáncer de cuello uterino y la posibilidad de presentar verrugas anogenitales. Se presenta información sobre estrategias de acciones de prevención y control, las cuales son importantes para entender el problema, ofrecer asistencia de calidad y tratamiento efectivo.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis / Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: pruebas de diagnóstico de sífilis

Resumo As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

Abstract The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Resumen Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.