Development and assessment of a multiepitope synthetic antigen for the diagnosis of Dengue virus infection.

Immunodiagnostic tests for detecting dengue virus infections encounter challenges related to cross-reactivity with other related flaviviruses. Our research focuses on the development of a synthetic multiepitope antigen tailored for dengue immunodiagnostics. Selected dengue epitopes involved structural linearity and dissimilarity from the proteomes of Zika and Yellow fever viruses which served for computationally modeling the three-dimensional protein structure, resulting in the design of two proteins: rDME-C and rDME-BR. Both proteins consist of seven epitopes, separated by the GPGPG linker, and a carboxy-terminal 6 × -histidine tag. The molecular weights of the final proteins rDME-C and rDME-BR are 16.83 kDa and 16.80 kDa, respectively, both with an isoelectric point of 6.35. The distinguishing factor between the two proteins lies in the origin of their epitope sequences, where rDME-C is based on the reference dengue proteome, while rDME-BR utilizes sequences from prevalent Dengue genotypes in Brazil from 2008 to 2019. PyMol analysis revealed exposure of epitopes in the secondary structure. Successful expression of the antigens was achieved in soluble form and fluorescence experiments indicated a disordered structure. In subsequent testing, rDME-BR and rDME-C antigens were assessed using an indirect Elisa protocol against Dengue infected serum, previously examined with a commercial diagnostic test. Optimal concentrations for antigens were determined at 10 µg/mL for rDME-BR and 30 µg/mL for rDME-C, with serum dilutions ranging from 1:50 to 1:100. Both antigens effectively detected IgM and IgG antibodies in Dengue fever patients, with rDME-BR exhibiting higher sensitivity. Our in-house test showed a sensitivity of 77.3 % and 82.6 % and a specificity of 89.4 % and 71.4 % for rDME-C and rDEM-BR antigens. No cross-reactivity was observed with serum from Zika-infected mice but with COVID-19 serum samples. Our findings underscore the utility of synthetic biology in crafting Dengue-specific multiepitope proteins and hold promise for precise clinical diagnosis and monitoring responses to emerging Dengue vaccines.

Lessons from a Multilaboratorial Task Force for Diagnosis of a Fatal Toxoplasmosis Outbreak in Captive Primates in Brazil.

Toxoplasmosis is an important zoonotic disease caused by the parasite Toxoplasma gondii and is especially fatal for neotropical primates. In Brazil, the Ministry of Health is responsible for national epizootic surveillance, but some diseases are still neglected. Here, we present an integrated investigation of an outbreak that occurred during the first year of the COVID-19 pandemic among eleven neotropical primates housed at a primatology center in Brazil. After presenting non-specific clinical signs, all animals died within four days. A wide range of pathogens were evaluated, and we successfully identified T. gondii as the causative agent within four days after necropsies. The liver was the most affected organ, presenting hemorrhage and hepatocellular necrosis. Tachyzoites and bradyzoite cysts were observed in histological examinations and immunohistochemistry in different organs; in addition, parasitic DNA was detected through PCR in blood samples from all specimens evaluated. A high prevalence of Escherichia coli was also observed, indicating sepsis. This case highlights some of the obstacles faced by the current Brazilian surveillance system. A diagnosis was obtained through the integrated action of researchers since investigation for toxoplasmosis is currently absent in national guidelines. An interdisciplinary investigation could be a possible model for future epizootic investigations in animals.

Case report: Regression of Glioblastoma after flavivirus infection.

Glioblastoma is the most frequent and aggressive primary brain cancer. In preclinical studies, Zika virus, a flavivirus that triggers the death of glioblastoma stem-like cells. However, the flavivirus oncolytic activity has not been demonstrated in human patients. Here we report a glioblastoma patient who received the standard of care therapy, including surgical resection, radiotherapy and temozolomide. However, shortly after the tumor mass resection, the patient was clinically diagnosed with a typical arbovirus-like infection, during a Zika virus outbreak in Brazil. Following the infection resolution, the glioblastoma regressed, and no recurrence was observed. This clinical response continues 6 years after the glioblastoma initial diagnosis.

Molecular mimicry between Zika virus and central nervous system inflammatory demyelinating disorders: the role of NS5 Zika virus epitope and PLP autoantigens / Mimetismo molecular entre o vírus Zika e os distúrbios inflamatórios desmielinizantes do sistema nervoso central: o papel do epítopo NS5 do vírus Zika e dos autoantígenos PLP

Abstract Background Evidence indicates a strong link between Zika virus (ZikV) and neurological complications. Acute myelitis, optic neuritis, polyneuropathy, and encephalomyelitis that mimic inflammatory idiopathic demyelination disorders (HDD) after ZikV infection have been reported in Brazil. Objective The present study aims to investigate the possible occurrence of molecular mimicry between ZikV antigens and Multiple Sclerosis (MS) autoantigens, the most frequent HDD of the central nervous system (CNS). Methods A retrospective cohort study with 305 patients admitted due to suspected arbovirus infection in Rio de Janeiro was performed, all subjects were submitted to neurological examination, and a biological sample was collected for serologic and molecular diagnostic. Bioinformatics tools were used to analyze the peptides shared between ZikV antigens and MS autoantigens. Results Of 305 patients, twenty-six were positive for ZikV and 4 presented IDD patterns found in MS cases. Sequence homology comparisons by bioinformatics approach between NS5 ZikV and PLP MS protein revealed a homology of 5/6 consecutive amino acids (CSSVPV/CSAVPV) with 83% identity, deducing a molecular mimicry. Analysis of the 3D structures revealed a similar conformation with alpha helix presentation. Conclusions Molecular mimicry between NS5 Zika virus antigen and PLP MS autoantigens emerge as a possible mechanism for IDD spectrum in genetically susceptible individuals.

Resumo Antecedentes Evidências indicam uma forte ligação entre o vírus Zika (ZikV) e complicações neurológicas. Mielite aguda, neurite óptica, polineuropatia e encefalomielite que mimetizam distúrbios inflamatórios de desmielinização idiopáticos (DDII) após infecção por ZikV têm sido relatadas no Brasil. Obejtivo O presente estudo tem como objetivo investigar a possível ocorrência de mimetismo molecular entre antígenos do ZikV e autoantígenos da Esclerose Múltipla (EM), a DDII mais frequente do sistema nervoso central (SNC). Métodos Foi realizado um estudo de coorte retrospectivo com 305 pacientes internados por suspeita de infecção por arbovirus no Rio de Janeiro, todos os indivíduos foram submetidos a exame neurológico e coleta de amostra biológica para diagnóstico sorológico e molecular. Ferramentas de bioinformática foram usadas para analisar os peptídeos compartilhados entre antígenos do ZikV e autoantígenos da EM. Resultados Dos 305 pacientes, vinte e seis foram positivos para ZikV e 4 apresentaram padrão IDD encontrado em casos de EM. As comparações de homologia de sequência por abordagem de bioinformática entre a proteína NS5 ZikV e PLP EM revelaram uma homologia de 5/6 aminoácidos consecutivos (CSSVPV/CSAVPV) com 83% de identidade, deduzindo um mimetismo molecular. A análise das estruturas 3D revelou uma conformação semelhante com apresentação em alfa-hélice. Conclusões O mimetismo molecular entre o antígeno NS5 do vírus Zika e o autoantígeno PLP da EM surge como um possível mecanismo para o espectro IDD em indivíduos geneticamente suscetíveis.

Identification and characterisation of SARS-CoV-2 and Human alphaherpesvirus 1 from a productive coinfection in a fatal COVID-19 case.

BACKGROUND: During routine Coronavirus disease 2019 (COVID-19) diagnosis, an unusually high viral load was detected by reverse transcription real-time polymerase chain reaction (RT-qPCR) in a nasopharyngeal swab sample collected from a patient with respiratory and neurological symptoms who rapidly succumbed to the disease. Therefore we sought to characterise the infection. OBJECTIVES: We aimed to determine and characterise the etiological agent responsible for the poor outcome. METHODS: Classical virological methods, such as plaque assay and plaque reduction neutralisation test combined with amplicon-based sequencing, as well as a viral metagenomic approach, were performed to characterise the etiological agents of the infection. FINDINGS: Plaque assay revealed two distinct plaque phenotypes, suggesting either the presence of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains or a productive coinfection of two different species of virus. Amplicon-based sequencing did not support the presence of any SARS-CoV-2 genetic variants that would explain the high viral load and suggested the presence of a single SARS-CoV-2 strain. Nonetheless, the viral metagenomic analysis revealed that Coronaviridae and Herpesviridae were the predominant virus families within the sample. This finding was confirmed by a plaque reduction neutralisation test and PCR. MAIN CONCLUSIONS: We characterised a productive coinfection of SARS-CoV-2 and Herpes simplex virus 1 (HSV-1) in a patient with severe symptoms that succumbed to the disease. Although we cannot establish the causal relationship between the coinfection and the severity of the clinical case, this work serves as a warning for future studies focused on the interplay between SARS-CoV-2 and HSV-1 coinfection and COVID-19 severity.

The incidence and geographical spread of SARS-CoV-2 in Rio de Janeiro, Brazil based on RT-PCR test results.

INTRODUCTION: Rio de Janeiro has hardly experienced coronavirus disease. METHODS: Here, 87,442 reverse transcription polymerase chain reaction (RT-PCR) test results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were reported among Rio de Janeiro residents (March to September 2020). RESULTS: Overall, RT-PCR positivity of 44.6% decreased over time towards 20%. Positivity was greater among males (OR=1.22; 95%CI:1.19-1.26); Black (OR=1.10; 95%CI:1.02-1.19), Brown (OR=1.16; 95%CI:1.10-1.22), and indigenous people (OR=2.11; 95%CI:0.88-5.03) compared to Whites and increased with age; with epidemic spread from the capital to inland regions. CONCLUSIONS: SARS-CoV-2 keeps spreading in Rio de Janeiro, and reopening of activities may fuel the epidemic.

Identification and characterisation of SARS-CoV-2 and Human alphaherpesvirus 1 from a productive coinfection in a fatal COVID-19 case

BACKGROUND During routine Coronavirus disease 2019 (COVID-19) diagnosis, an unusually high viral load was detected by reverse transcription real-time polymerase chain reaction (RT-qPCR) in a nasopharyngeal swab sample collected from a patient with respiratory and neurological symptoms who rapidly succumbed to the disease. Therefore we sought to characterise the infection. OBJECTIVES We aimed to determine and characterise the etiological agent responsible for the poor outcome. METHODS Classical virological methods, such as plaque assay and plaque reduction neutralisation test combined with amplicon-based sequencing, as well as a viral metagenomic approach, were performed to characterise the etiological agents of the infection. FINDINGS Plaque assay revealed two distinct plaque phenotypes, suggesting either the presence of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains or a productive coinfection of two different species of virus. Amplicon-based sequencing did not support the presence of any SARS-CoV-2 genetic variants that would explain the high viral load and suggested the presence of a single SARS-CoV-2 strain. Nonetheless, the viral metagenomic analysis revealed that Coronaviridae and Herpesviridae were the predominant virus families within the sample. This finding was confirmed by a plaque reduction neutralisation test and PCR. MAIN CONCLUSIONS We characterised a productive coinfection of SARS-CoV-2 and Herpes simplex virus 1 (HSV-1) in a patient with severe symptoms that succumbed to the disease. Although we cannot establish the causal relationship between the coinfection and the severity of the clinical case, this work serves as a warning for future studies focused on the interplay between SARS-CoV-2 and HSV-1 coinfection and COVID-19 severity.

The incidence and geographical spread of SARS-CoV-2 in Rio de Janeiro, Brazil based on RT-PCR test results

Abstract INTRODUCTION Rio de Janeiro has hardly experienced coronavirus disease. METHODS Here, 87,442 reverse transcription polymerase chain reaction (RT-PCR) test results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were reported among Rio de Janeiro residents (March to September 2020). RESULTS Overall, RT-PCR positivity of 44.6% decreased over time towards 20%. Positivity was greater among males (OR=1.22; 95%CI:1.19-1.26); Black (OR=1.10; 95%CI:1.02-1.19), Brown (OR=1.16; 95%CI:1.10-1.22), and indigenous people (OR=2.11; 95%CI:0.88-5.03) compared to Whites and increased with age; with epidemic spread from the capital to inland regions. CONCLUSIONS SARS-CoV-2 keeps spreading in Rio de Janeiro, and reopening of activities may fuel the epidemic.

Laboratory Acquired Zika Virus Infection Through Mouse Bite: A Case Report.

Zika virus is an arthropod-borne flavivirus mainly transmitted by the bite of infected mosquitoes. However, alternative transmission routes can occur. In this study, we show the accidental transmission of virus from an infected mouse to a human during the experimental manipulation. This study describes the patient clinical manifestations and virus genome identification.

Application of the Respondent-Driven Sampling methodology in a biological and behavioral surveillance survey among female sex workers, Brazil, 2016.

INTRODUCTION: Respondent-Driven Sampling (RDS) has been used in surveys with key populations at risk of HIV infection, such as female sex workers (FSW). This article describes the application of the RDS method among FSW in 12 Brazilian cities, during a survey carried out in 2016. METHODOLOGY: A biological and behavioral surveillance study carried out in 12 Brazilian cities, with a minimum sample of 350 FSW in each city. Tests were performed for HIV, syphilis, and hepatitis B and C infections. A social-behavioral questionnaire was also applied. RESULTS: The sample was comprised of 4,328 FSW. For data analysis, the sample was weighted according to each participant's network size (due consideration to the implications of RDS complex design and to the effects of homophilia are recommended). DISCUSSION: Although RDS methods for obtaining a statistical sample are based on strong statistical assumptions, allowing for an estimation of statistical parameters, with each new application the method has been rethought. In the analysis of whole-sample data, estimators were robust and compatible with those found in 2009. However, there were significant variations according to each city. CONCLUSION: The achieved sample size was of great relevance for assessing progress and identifying problems regarding the prevention and treatment of Sexually Transmitted Infections. New RDS studies with more time and operational resources should be envisaged. This could further network development.

Application of the Respondent-Driven Sampling methodology in a biological and behavioral surveillance survey among female sex workers, Brazil, 2016 / Aplicação da metodologia Respondent-Driven Sampling em pesquisa biológica e comportamental com mulheres trabalhadoras do sexo, Brasil, 2016

ABSTRACT Introduction: Respondent-Driven Sampling (RDS) has been used in surveys with key populations at risk of HIV infection, such as female sex workers (FSW). This article describes the application of the RDS method among FSW in 12 Brazilian cities, during a survey carried out in 2016. Methodology: A biological and behavioral surveillance study carried out in 12 Brazilian cities, with a minimum sample of 350 FSW in each city. Tests were performed for HIV, syphilis, and hepatitis B and C infections. A social-behavioral questionnaire was also applied. Results: The sample was comprised of 4,328 FSW. For data analysis, the sample was weighted according to each participant's network size (due consideration to the implications of RDS complex design and to the effects of homophilia are recommended). Discussion: Although RDS methods for obtaining a statistical sample are based on strong statistical assumptions, allowing for an estimation of statistical parameters, with each new application the method has been rethought. In the analysis of whole-sample data, estimators were robust and compatible with those found in 2009. However, there were significant variations according to each city. Conclusion: The achieved sample size was of great relevance for assessing progress and identifying problems regarding the prevention and treatment of Sexually Transmitted Infections. New RDS studies with more time and operational resources should be envisaged. This could further network development.

RESUMO Introdução: O método de amostragem Respondent-Driven Sampling (RDS) tem sido utilizado em inquéritos com populações-chave sob maior risco de infecção pelo HIV, como as mulheres trabalhadoras do sexo (MTS). Este artigo tem o objetivo de descrever a implementação do RDS entre MTS em 12 cidades brasileiras em 2016. Metodologia: Trata-se de um estudo de vigilância biológica e comportamental realizado em 12 cidades brasileiras, com amostra mínima de 350 MTS em cada cidade. Foram realizados testes para infecções por HIV, sífilis, hepatites B e C, e aplicou-se questionário sociocomportamental. Resultados: Participaram 4.328 MTS. Para a análise dos dados, foi elaborada uma ponderação amostral considerando o tamanho da rede de cada participante; recomenda-se que o desenho complexo de amostragem por RDS e o efeito de homofilia sejam considerados. Discussão: Apesar de o RDS ser fundamentado em pressupostos estatísticos para obtenção de uma amostra probabilística e possibilitar estimação de parâmetros estatísticos, ele vem sendo repensado a cada nova aplicação. Na análise dos dados na totalidade da amostra, os estimadores mostraram-se robustos e coerentes aos encontrados em 2009. Entretanto, constataram-se grandes variações por cidade. Conclusão: O tamanho amostral alcançado foi de grande relevância para avaliar avanços e identificar problemas a respeito da prevenção e assistência às infecções sexualmente transmissíveis. Ressalta-se a necessidade de pensar estudos RDS com maior tempo e recursos para implementação, o que poderia permitir um melhor desenvolvimento das redes.

The influence of combat simulation exercises on indirect markers of muscle damage in soldiers of the brazilian army / A influência do exercício de simulação de combate sobre marcadores indiretos de lesão muscular em soldados do exército brasileiro

The combat simulation exercise is a military activity where combat activities characterized by intense physical activity are simulated. The aim of this study was to describe the behavior of indirect markers of muscle damage during military activities of combat simulation with the realization of strenuous physical efforts. This study was conducted with military recruits/volunteers (n=43; 19-24 age group) from the COMANDOS Army Corporal Training Course. The biomarkers were evaluated at the baseline (T0), 72 hours after the baseline (T1) and 63 hours after the end of the activity (T2). To compare the variables analyzed at different times, an analysis of variance (ANOVA) was applied to repeated measures with post hoc Tukey's test to identify the statistically significant differences. The criteria of significance adopted was the value of p<0,05. On average, the total body mass was significantly greater in T0 when compared to T1 (74,4kg x 69,8Kg; p< 0,05). In relation to the biochemical markers, on average, the serum levels were significantly higher (p<0,001) in T1 when compared to T0: CPK, LDH, CRP, Mb and AGPA. After 63 hours of rest, on average, the serum levels of the biochemical markers were significantly lower than in the moment T1: CPK, LDH, CRP, Mb and AGPA. However, after 63 hours of the end of the activity, all variables evaluated returned to baseline, with the exception of LDH, that on an average remained above the values observed in the moment T0. This study made possible to understand the behavior of indirect markers of muscle damage during the COMANDOS Army Corporal Training Course. However, others studies are necessary, regarding to Training Courses such as: paratrooper, mountain, jungle warfare and others to try establish a possible range of reference values for this markers during military activities.

O exercício de simulação de combate é uma atividade militar, onde as atividades de combate caracterizadas por intensa atividade física são simuladas. O objetivo deste estudo foi descrever o comportamento dos marcadores indiretos de lesão muscular durante as atividades militares de simulação de combate com a realização de esforços físicos extenuantes. Este estudo foi realizado com militares voluntários (n = 43; faixa etária = 19-24 anos) do Curso de Formação de Cabos do Exército. Os biomarcadores foram avaliados na linha de base (T0), 72 horas após o início das atividades (T1) e 63 horas após o final da atividade de (T2). Uma análise de variância (ANOVA) para medidas repetidas, com teste post hoc Tukey, foi aplicada para identificar as diferenças estatisticamente significativas nas variáveis analisadas em momentos diferentes. O critério de significância adotado foi o valor de p <0,05. Em média, a massa corporal total foi significativamente maior no T0 quando comparado a T1 (74,4kg x 69,8Kg; p <0,05). Em relação aos marcadores bioquímicos, em média, os níveis séricos foram significativamente maiores (p <0,001) em T1 quando comparados com T0 para: CPK, LDH, CRP, Mb e AGPA. Após 63 horas de descanso, em média, os níveis séricos dos marcadores bioquímicos (CPK, LDH, CRP, Mb e AGPA ) foram significativamente menores do que no momento T1. No entanto, após 63 horas a partir do fim da atividade, todas as variáveis avaliadas retornaram à linha de base, com a exceção do LDH, que, em média, manteve-se significativamente maior do que os valores observados no momento T0. Este estudo possibilitou compreender o comportamento dos marcadores indiretos de lesão muscular durante o Curso de Formação de Cabos COMANDOS. No entanto, outros estudos são necessários, com relação a cursos de formação, tais como: paraquedistas, montanha, guerra na selva, e outros, para tentar estabelecer uma possível faixa de valores de referência para estes marcadores durante as atividades militares.

Changes in attitudes, risky practices, and HIV and syphilis prevalence among female sex workers in Brazil from 2009 to 2016.

BACKGROUND: The present study aims at investigating the progress made toward controlling the human immunodeficiency virus (HIV)/AIDS epidemic among female sex workers (FSW) from 2009 to 2016. METHODS: The baseline of respondent-driven sampling (RDS) study among FSW was carried out in 2009, in 10 Brazilian municipalities. In 2016, information on FSW were collected in 12 municipalities. The analyses took into account the dependence among observations, resulting from the recruitment chains, and the unequal probabilities of selection, resulting from the different network sizes. We analyzed changes in attitudes and risky behavior practices as well as variations in HIV and syphilis prevalence based on the comparison of 95% confidence intervals for each estimate. RESULTS: Information on 2523 (2009) and 4245 (2016) FSW were analyzed. Commercial sex debut shifted to younger ages: while in 2009 the proportion of women who started sex work under 18 years old was 28.3%, in 2016 this percentage rose to 38.3%. The proportion of FSW affiliated to a nongovernmental organization (NGO) in defense of their rights (14.0%), in 2009, decreased to 7.8%, in 2016, as well as the proportion of FSW who received counseling on sexually transmitted infections (STI) in the past 6 months, from 47.5% to 24.4%. Relevant improvements were found for HIV testing in the last 12 months (from 20.3% to 39.3%). The proportions of those who were never tested for syphilis dropped from 57.9% to 48.5%. However, an opposite decreasing trend was found for the Pap smear examination in the last 12 months, decreasing from 43.6% to 31.5%. Regular condom use with clients significantly increased in the period. Regarding HIV prevalence, the 5% level was sustained and no significant differences were found, but syphilis prevalence was found to be more than 3 times higher in 2016 (8.5%) than in 2009 (2.4%). DISCUSSION: Many are the challenges to be faced in attempting to reverse the upward trend of syphilis among FSW in Brazil. Despite the progress in condom distribution free of charge, it is necessary to increase awareness campaigns, emphasize the use, reaffirm STI counseling, and reiterate the need of regular syphilis screening in this key population group.

Factors associated with HIV infection among female sex workers in Brazil.

BACKGROUND: Female sex workers (FSWs) are one of the most-at-risk population groups for human immunodeficiency virus (HIV) infection. This paper aims at identifying the main predictors of HIV infection among FSW recruited in the 2nd Biological and Behavioral Surveillance Survey in 12 Brazilian cities in 2016. METHOD: Data were collected on 4245 FSW recruited by respondent driven sampling (RDS). Weights were inversely proportional to participants' network sizes. To establish the correlates of HIV infection, we used logistic regression models taking into account the dependence of observations resultant from the recruitment chains. The analysis included socio-demographic sex work characteristics, sexual behavior, history of violence, alcohol and drug use, utilization of health services, and occurrence of other sexually transmitted infections (STIs). RESULTS: HIV prevalence was estimated as 5.3% (4.4%-6.2%). The odds ratio (OR) of an HIV-positive recruiter choosing an HIV-positive participant was 3.9 times higher than that of an HIV-negative recruiter (P < .001). Regarding socio-demographic and sex work characteristics, low educational level, street as the main work venue, low price per sexual encounter, and longer exposure time as a sex worker were found to be associated with HIV infection, even after controlling for the homophily effect. The OR of being HIV infected among FSW who had been exposed to sexual violence at least once in a lifetime (OR = 1.5, P = .028) and the use of illicit drugs at least once a week were highly significant as well, particularly for frequent crack use (OR = 3.6, P < .001). Among the sexual behavior indicators, not using condoms in some circumstances were significantly associated with HIV infection (OR = 1.8, P = .016). Regarding the occurrence of other STI, the odds of being HIV infected was significantly higher among FSW with a reactive treponemal test for syphilis (OR = 4.6, P < .001). CONCLUSIONS: The main factors associated with HIV infection identified in our study characterize a specific type of street-based sex work in Brazil and provided valuable information for developing interventions. However, there is a further need of addressing social and contextual factors, including illicit drug use, violence, exploitation, as well as stigma and discrimination, which can influence sexual behavior.

Prevalence estimates of HIV, syphilis, hepatitis B and C among female sex workers (FSW) in Brazil, 2016.

BACKGROUND: Female sex workers (FSW) bear a high burden of sexually transmitted infections (STI). In this paper, we estimate the prevalence of human immunodeficiency virus (HIV), HBV = hepatitis B virus (HBV), HCV = hepatitis C virus (HCV), syphilis and co-infections in the second Biological and Behavioral Surveillance Survey among FSW in Brazil. METHOD: The survey was conducted in 12 Brazilian cities from July to November 2016. We used respondent-driven sampling (RDS) to recruit 350 FSW in each city. Rapid tests were used for screening HIV, syphilis, HCV, and HBV. Confirmatory tests were performed on all samples with reactive rapid test result. All testing algorithms and interpretations were done according to the recommendations of the Department of STI/AIDS and viral hepatitis, Ministry of Health. The STI diagnoses were given by: confirmed HIV infection by a positive result on Western blot; active syphilis infection, defined by a RPR titer equal or greater than 1/8; viremia period of HBV and HCV infections, characterized by a detectable (or quantifiable) viral load. Prevalence estimates and standard errors were calculated using statistical procedures suitable for data collected by RDS. RESULTS: Excluding the seeds, 4245 FSW were enrolled. Prevalence estimates were: HIV 5.3% (95% CI: 4.4%-6.2%); active syphilis 8.5% (95% CI: 7.3%-9.7%); HBV 0.4% (95% CI: 0.2%-0.7%); and, HCV 0.9% (95% CI: 0.6%-1.3%). Among the 4154 FSW tested for the 4 infections, 13.3%; (95% CI 12.0%-14.8%) were diagnosed with at least one of the infections, of which 87.6% (95% CI: 83.3%-90.9%) had single infections. The prevalence of HIV/syphilis co-infection was 1.09% (95% CI: 0.7%- 1.6%) and of HIV/HCV or HBV infections was 0.4% (95% CI: 0.2%-0.7%). CONCLUSIONS: Our results reveal the need to conduct more studies to estimate the prevalence of STI and co-infections among FSW in Brazil. Longitudinal trends in the prevalence estimates of HIV and other STI provide information to monitor changes in this high-risk population. Additionally, the study highlights the importance of measuring the hepatitis burden among FSW living with HIV, and the need of including FSW in all aspects of STI prevention, care, and treatment programs.

Evaluation of seven rapid tests for syphilis available in Brazil using defibrinated plasma panels.

OBJECTIVES: In 2012, the WHO estimated that 6 million new cases of syphilis per year would occur worldwide, including 937 000 in Brazil. Early diagnosis and treatment of syphilis are essential to reduce morbidity and prevent transmission. The availability of rapid tests (RTs) for this diagnosis means that testing can be performed more quickly, as a point-of-care test, even in non-laboratory environments and requires only simple technical training to antibodies detection. The objective of this study was to evaluate the performance and operational aspects of seven commercially available RTs for syphilis in Brazil. METHODS: Seven rapid treponemal tests were evaluated for sensitivity, specificity, accuracy and Kappa value, according to a panel composed of 493 members. The operational performance of the assay was also determined for these tests. RESULTS: The seven RTs showed sensitivity ranging from 94.5% to 100% when compared with the reference tests and specificity of between 91.5% and 100%. All the RTs evaluated presented good operational performance, and only one failed to present the minimum specificity as defined by Brazil's Ministry of Health. CONCLUSION: All the tests presented good operational performance, and the professionals who performed them considered them to be easy to use and interpret. This evaluation is important for making informed choices of tests to be used in the Brazilian Unified Health System.

Estimation of HIV incidence in two Brazilian municipalities, 2013 / Estimação da incidência de HIV em dois municípios brasileiros, 2013

ABSTRACT OBJECTIVE To estimate HIV incidence in two Brazilian municipalities, Recife and Curitiba, in the year of 2013. METHODS The method for estimating incidence was based on primary information, resulting from the Lag-Avidity laboratory test for detection of recent HIV infections, applied in a sample of the cases diagnosed in the two cities in 2013. For the estimation of the HIV incidence for the total population of the cities, the recent infections detected in the research were annualized and weighted by the inverse of the probability of HIV testing in 2013 among the infected and not diagnosed cases. After estimating HIV incidence for the total population, the incidence rates were estimated by sex, age group, and exposure category. RESULTS In Recife, 902 individuals aged 13 years and older were diagnosed with HIV infection. From these, 528 were included in the study, and the estimated proportion of recent infections was 13.1%. In Curitiba, 1,013 people aged 13 years and older were diagnosed, 497 participated in the study, and the proportion of recent infections was 10.5%. In Recife, the estimated incidence rate was 53.1/100,000 inhabitants of 13 years and older, while in Curitiba, it was 41.1/100,000, with male-to-female ratio of 3.5 and 2.4, respectively. We observed high rates of HIV incidence among men who have sex with men, of 1.47% in Recife and 0.92% in Curitiba. CONCLUSIONS The results obtained in the two cities showed that the group of men who have sex with men are disproportionately subject to a greater risk of new infections, and indicate that strategies to control the spread of the epidemic in this population subgroup are essential and urgent.

RESUMO OBJETIVO Estimar a incidência de HIV em dois municípios brasileiros, Recife e Curitiba, no ano de 2013. MÉTODOS O método de estimação da incidência foi baseado em informações primárias, resultantes do ensaio laboratorial Lag-Avidity para detecção de infecções recentes do HIV, aplicado em uma amostra dos casos diagnosticados nas duas cidades em 2013. Para a estimação da incidência de HIV para a população total das cidades, as infecções recentes detectadas na pesquisa foram anualizadas e ponderadas pelo inverso da probabilidade de teste de HIV no ano de 2013 entre os casos infectados e não diagnosticados. Após a estimação da incidência de HIV para a população total, foram estimadas as taxas de incidência por sexo, faixa de idade e categoria de exposição. RESULTADOS Em Recife, foram diagnosticados 902 indivíduos de 13 anos e mais com infecção de HIV. Desses, 528 foram incluídos no estudo, e a proporção estimada de infecções recentes foi de 13,1%. Em Curitiba, foram diagnosticadas 1.013 pessoas de 13 anos e mais, 497 participaram do estudo, e a proporção de infecções recentes foi de 10,5%. Em Recife, a taxa de incidência estimada foi de 53,1 por 100 mil habitantes de 13 anos e mais, enquanto em Curitiba, de 41,1 por 100 mil, com razão do sexo masculino para o feminino de 3,5 e 2,4, respectivamente. Foram evidenciadas elevadas taxas de incidência de HIV entre homens que fazem sexo com homens, de 1,47% em Recife e 0,92% em Curitiba. CONCLUSÕES Os resultados obtidos nas duas cidades mostraram que o grupo dos homens que fazem sexo com homens está desproporcionalmente sujeito ao maior risco de novas infecções, e indicam que estratégias para controle da disseminação da epidemia nesse subgrupo populacional são essenciais e urgentes.

Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20 percent. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24 percent) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients.

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20%. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24%) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

Evaluation of three methods for hemoglobin. Measurement in a blood donor setting

Context: The hemoglobin (HB) level is the most-used parameter for screening blood donors for the presence of anemia, one of the most-used methods for measuring Hb levels is based on photometric detection of cyanmetahemoglobin, as an alternative to this technology, Hemo Cue has developed a photometric method based on the determination of azide metahemoglobin. Objective: To evaluate the performance of three methods for hemoglobin (Hb) determination in a blood bank setting. Design: Prospective study utilizing samples to compare methods for Hb determination. Setting: Hemotherapy Service of the Hospital Israelita Albert Eistein, a private institution in the tertiary health care system. Samples: Seria blood samples were collected from 259 individuals during the period from March to June 1996. Main Measurements: Test performances and their comparisions were assessed by the analysis of coefficients of variation (CV), linear regression and mean differences. Results: The CV for the three methods were: Coulter 0.68 per cent, Cobas 0.82 per cent and Hemo Cue 0.69 per cent. There was no difference between the mean Hb determination for the three methods (p>0.05). The Coulter and Cobas methods showed the best agreement and the HemoCue method gave a lower Hb determination when compared to both the Coulter and Cobas methods. However, paris of methods involving the Hemo Cue seem to have narrower limits of agreement (+0.78 and + 1.02) than the Coulter and Cobas combination (+ 1.13). Conclusion: The three methods provide good agreement for hemoglobin determination.