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Genomic surveillance has been applied since the beginning of the COVID-19 pandemic to track the spread of the virus, leading to the characterization of multiple SARS-CoV-2 variants, including variants of concern (VOC). Although sequencing is the standard method, a rapid molecular test for screening and surveillance of VOC is considered for detection. Furthermore, using alternative saliva as specimen collection facilitates the implementation of a less invasive, self-collected sample. In this study, we applied a combinatory strategy of saliva collection and reverse transcription polymerase chain reaction (RT-PCR) for SARS-CoV-2 VOC detection. Saliva samples from patients attending a tertiary hospital with suspected COVID-19 were collected and SARS-CoV-2 RNA was detected using SARS-CoV-2 RT-qPCR reagent kit (PerkinElmer). Positive saliva samples were screened for SARS-CoV-2 VOC with previously described RT-PCR for Alpha, Beta, and Gamma variants. Saliva samples were positive in 171 (53%) of 324 tested. A total of 108 (74%) from positive samples were also positive for VOC by RT-PCR screening. Those samples were found between January and August 2021. This approach allowed us to successfully use an alternative and complementary tool to genomic surveillance to monitor the circulation of SARS-CoV-2 VOC in the studied population.
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COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , COVID-19/epidemiologia , Humanos , Pandemias , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , SARS-CoV-2/genética , SalivaRESUMO
BACKGROUND Despite efforts to mitigate the impact of dengue virus (DENV) epidemics, the virus remains a public health problem in tropical and subtropical regions around the world. Most DENV cases in the Americas between January and July 2019 were reported in Brazil. São Paulo State in the southeast of Brazil has reported nearly half of all DENV infections in the country. OBJECTIVES To understand the origin and dynamics of the 2019 DENV outbreak. METHODS Here using portable nanopore sequencing we generated20 new DENV genome sequences from viremic patients with suspected dengue infection residing in two of the most-affected municipalities of São Paulo State, Araraquara and São José do Rio Preto. We conducted a comprehensive phylogenetic analysis with 1,630 global DENV strains to better understand the evolutionary history of the DENV lineages that currently circulate in the region. FINDINGS The new outbreak strains were classified as DENV2 genotype III (American/Asian genotype). Our analysis shows that the 2019 outbreak is the result of a novel DENV lineage that was recently introduced to Brazil from the Caribbean region. Dating phylogeographic analysis suggests that DENV2-III BR-4 was introduced to Brazil in or around early 2014, possibly from the Caribbean region. MAIN CONCLUSIONS Our study describes the early detection of a newly introduced and rapidly-expanding DENV2 virus lineage in Brazil.
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Vírus da Dengue/genética , Dengue/virologia , Variação Genética , Genômica , Brasil , Genótipo , Humanos , Filogenia , RNA Viral/genéticaRESUMO
Several countries have local transmission of multiple arboviruses, in particular, dengue and Zika viruses, which have recently spread through many American countries. Cross reactivity among Flaviviruses is high and present a challenge for accurate identification of the infecting agent. Thus, we evaluated the level of cross reactivity of anti-dengue IgM/G Enzyme-Linked Immunosorbent Assays (ELISA) from three manufacturers against 122 serum samples obtained at two time-points from 61 patients with non-dengue confirmed Zika virus infection. All anti-dengue ELISAs cross reacted with serum from patients with acute Zika infection at some level and a worrisome number of seroconversion for dengue IgG and IgM was observed. These findings may impact the interpretation of currently standard criteria for dengue diagnosis in endemic regions.
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Reações Cruzadas , Dengue/diagnóstico , Testes Diagnósticos de Rotina/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Infecção por Zika virus/diagnóstico , Adulto , Anticorpos Antivirais/sangue , Erros de Diagnóstico , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , VirosesRESUMO
Fever, skin rash, headache, and thrombocytopenia are considered hallmarks of dengue infection. However, these symptoms are frequently observed in infectious and non-infectious complications of hematopoietic stem cell transplant recipients and oncohematological patients. Thus, laboratory confirmation of dengue is relevant for prompt intervention and proper management of dengue in endemic and non-endemic regions. Because no prospective study of dengue has been conducted in these populations, the actual morbidity and mortality of dengue is unknown. In the present series, we describe five cases of dengue in patients living in endemic areas, emphasizing the prolonged course of the disease and the occurrence of prolonged viremia.
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Vírus da Dengue/isolamento & purificação , Dengue/diagnóstico , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Viremia/diagnóstico , Adolescente , Idoso , Pré-Escolar , Dengue/virologia , Vírus da Dengue/genética , Feminino , Febre , Neoplasias Hematológicas , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Trombocitopenia , Viremia/virologiaRESUMO
BACKGROUND: Dengue virus transmission by blood transfusion is a rarely reported event. CASE REPORT: During a dengue outbreak in São Paulo city, a regular plateletpheresis donor informed the blood bank of being diagnosed a few days after donation. The recipient was hospitalized and displayed symptoms and laboratory evidence of dengue after transfusion. RESULTS: The donor was immunoglobulin (Ig)G, IgM, and polymerase chain reaction nonreactive on the index sample, seroconverting 20 days later. The platelet units were transfused into two patients. One of them developed fever 3 days after transfusion, with high viral load. His pretransfusion sample was negative for IgG, IgM, and dengue RNA, while the second recipient did not show any symptoms nor laboratory evidence of dengue infection. CONCLUSIONS: This case brings additional evidence that dengue is indeed transmissible by blood transfusion and clinical manifestations, although rare, do occur.
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Dengue/etiologia , Dengue/transmissão , Reação Transfusional , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Here, we report application of high-throughput near full-length genome (NFLG) and partial human immunodeficiency virus Type 1 (HIV-1) proviral genome deep sequencing to characterize HIV in recently infected blood donors at four major blood centers in Brazil. STUDY DESIGN AND METHODS: From 2007 to 2011, a total of 341 HIV+ blood donors from four blood centers were recruited to participate in a case-control study to identify HIV risk factors and motivations to donate. Forty-seven (17 from São Paulo, eight from Minas Gerais, 11 from Pernambuco, and 11 from Rio de Janeiro) were classified as recently infected based on testing by less-sensitive enzyme immunoassays. Five overlapping amplicons spanning the HIV genome were polymerase chain reaction amplified from peripheral blood mononuclear cells. The amplicons were molecularly barcoded, pooled, and sequenced by a paired-end protocol (Illumina). RESULTS: Of the 47 recently infected donor samples studied, 39 (82.9%) NFLGs and six (12.7%) partial fragments were de novo assembled into contiguous sequences and successfully subtyped. Subtype B was the only nonrecombinant virus identified in this study and accounted for 62.2% (28/45) of samples. The remaining 37.8% (17/45) of samples showed various patterns of subtype discordance in different regions of HIV-1 genomes, indicating two to four circulating recombinant subtypes derived from Clades B, F, and C. Fourteen samples (31.1%) from this study harbored drug resistance mutations, indicating higher rate of drug resistance among Brazilian blood donors. CONCLUSION: Our findings revealed a high proportion of HIV-1 recombinants among recently infected blood donors in Brazil, which has implications for future blood screening, diagnosis, therapy, and vaccine development.
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Genoma Viral/genética , HIV-1/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Doadores de Sangue/estatística & dados numéricos , Brasil , Humanos , Técnicas Imunoenzimáticas , Dados de Sequência MolecularRESUMO
We conducted a spatial case-control study nested in a dengue incidence cohort to explore the role of the spatial and socioeconomic factors in the proportion of symptomatic (cases) and inapparent primary dengue virus infections (controls). Cohort participants were children and adolescents (2 to 16 years of age) at the beginning of the follow-up. Case definitions were, for symptomatic cases, fever plus a positive lab result for acute dengue (NS1, RT-PCR, ELISA IgM/IgG), and for inapparent infection a positive result for dengue IgG (ELISA) in subjects without symptoms and with a previously negative result at baseline. The covariates included sociodemographic factors, residential location, and socioeconomic context variables of the census tracts of residence of cases and controls. We used principal component analysis to reduce the contextual covariates, with the component values assigned to each one based on their residences. The data were modeled in a Bayesian context, considering the spatial dependence. The final sample consisted of 692 children, 274 cases and 418 controls, from the first year of follow-up (2014-2015). Being male, older age, higher educational level of the head of the family and having a larger number of rooms in the household were associated with a greater chance of presenting dengue symptomatic infection at the individual level. The contextual covariates were not associated with the outcome. Inapparent dengue infection has extensive epidemiological consequences. Relying solely on notifications of symptomatic dengue infections underestimates the number of cases, preserves a silent source of the disease, potentially spreading the virus to unaffected areas.
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Vírus da Dengue , Dengue , Criança , Adolescente , Humanos , Masculino , Feminino , Vírus da Dengue/genética , Dengue/diagnóstico , Dengue/epidemiologia , Estudos de Casos e Controles , Infecções Assintomáticas/epidemiologia , Brasil/epidemiologia , Teorema de Bayes , Imunoglobulina G , Anticorpos AntiviraisRESUMO
Dengue virus (DENV) is a prominent arbovirus with global spread, causing approximately 390 million infections each year. In Brazil, yearly epidemics follow a well-documented pattern of serotype replacement every three to four years on average. Araraquara, located in the state of São Paulo, has faced significant impacts from DENV epidemics since the emergence of DENV-1 in 2010. The municipality then transitioned from low to moderate endemicity in less than 10 years. Yet, there remains an insufficient understanding of virus circulation dynamics, particularly concerning DENV-1, in the region, as well as the genetic characteristics of the virus. To address this, we sequenced 37 complete or partial DENV-1 genomes sampled from 2015 to 2022 in Araraquara. Then, using also Brazilian and worldwide DENV-1 sequences we reconstructed the evolutionary history of DENV-1 in Araraquara and estimated the time to the most recent common ancestor (tMRCA) for serotype 1, for genotype V and its main lineages. Within the last ten years, there have been at least three introductions of genotype V in Araraquara, distributed in two main lineages (L Ia and L Ib, and L II). The tMRCA for the first sampled lineage (2015/2016 epidemics) was approximately 15 years ago (in 2008). Crucially, our analysis challenges existing assumptions regarding the emergence time of the DENV-1 genotypes, suggesting that genotype V might have diverged more recently than previously described. The presence of the two lineages of genotype V in the municipality might have contributed to the extended persistence of DENV-1 in the region.
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Vírus da Dengue , Dengue , Humanos , Filogenia , Vírus da Dengue/genética , Dengue/epidemiologia , Brasil/epidemiologia , GenótipoRESUMO
OBJECTIVES: The aim of this study was to achieve greater specificity of dengue virus (DENV) serological tests based on a recombinant antigen derived from non-structural protein 1 (ΔNS1) with regard to cross-reactive Zika virus (ZIKV) anti-NS1 antibody responses. This is of relevance in endemic regions for the serological discrimination of both DENV and ZIKV, such as Brazil and other tropical countries. METHODS: The ΔNS1 proteins were obtained as recombinant antigens and were evaluated as solid-phase-bound antigens in the ELISA test to detect anti-NS1 IgG antibodies. The performance of the ∆NS1-based DENV IgG ELISA was assessed with both mouse and human serum samples previously exposed to DENV or ZIKV. RESULTS: The ∆NS1-based DENV IgG ELISA detected anti-DENV NS1 IgG without cross-reactivity with ZIKV-positive serum samples. The sensitivity and specificity of the assay determined using samples previously characterized by real-time PCR (qRT-PCR) or plaque reduction neutralization assay (PRNT) were 82% and 93%, respectively. CONCLUSION: The ∆NS1-based DENV IgG ELISA conferred enhanced diagnostic specificity for anti-DENV serological tests and may be particularly useful for serological analyses in endemic regions for both DENV and ZIKV transmission.
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Vírus da Dengue , Dengue , Infecção por Zika virus , Zika virus , Animais , Anticorpos Antivirais , Dengue/diagnóstico , Ensaio de Imunoadsorção Enzimática , Camundongos , Sensibilidade e Especificidade , Proteínas não Estruturais Virais , Infecção por Zika virus/diagnósticoRESUMO
Reinfection by the severe acute respiratory syndrome coronavirus type 2 (SARS-COV-2) has been reported in many countries, suggesting that the virus may continue to circulate among humans despite the possibility of local herd immunity due to massive previous infections. The emergence of variants of concern (VOC) that are more transmissible than the previous circulating ones has raised particular concerns on the vaccines effectiveness and reinfection rates. The P.1 lineage was first identified in December 2020 in Manaus city and is now globally spread. We report the first case of reinfection of SARS-CoV-2 caused by the P.1 variant outside of Manaus. The potential of these new variants to escape naturally and vaccine- induced immunity highlights the need for a global vigilance.
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COVID-19 , Reinfecção , SARS-CoV-2 , Brasil/epidemiologia , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/virologia , Humanos , Reinfecção/virologia , SARS-CoV-2/isolamento & purificaçãoRESUMO
Background: Previous reported neurologic sequelae associated with SARS-CoV-2 infection have mainly been confined to hospital-based patients in which viral detection was restricted to nasal/throat swabs or to IgM/IgG peripheral blood serology. Here we describe seven cases from Brazil of outpatients with previous mild or moderate COVID-19 who developed subacute cognitive disturbances. Methods: From June 1 to August 15, 2020, seven individuals 18 to 60 years old, with confirmed mild/moderate COVID-19 and findings consistent with encephalopathy who were observed >7 days after respiratory symptom initiation, were screened for cognitive dysfunction. Paired sera and CSF were tested for SARS-CoV-2 (IgA, IgG ELISA, and RT-PCR). Serum and intrathecal antibody dynamics were evaluated with oligoclonal bands and IgG index. Cognitive dysfunction was assessed by the Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and the Clock Drawing Test (CDT). Results: All but one of our patients were female, and the mean age was 42.6 years. Neurologic symptoms were first reported a median of 16 days (IQR 15-33) after initial COVID-19 symptoms. All patients had headache and altered behavior. Cognitive dysfunction was observed mainly in phonemic verbal fluency (MoCA) with a median of six words/min (IQR 5.25-10.75) and altered visuospatial construction with a median of four points (IQR 4-9) (CDT). CSF pleocytosis was not detected, and only one patient was positive for SARS-Co Conclusions: A subacute cognitive syndrome suggestive of SARS-CoV-2-initiated damage to cortico-subcortical associative pathways that could not be attributed solely to inflammation and hypoxia was present in seven individuals with mild/moderate COVID-19.
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Intrahost genetic diversity is thought to facilitate arbovirus adaptation to changing environments and hosts, and it might also be linked to viral pathogenesis. Dengue virus serotype 2 (DENV-2) has circulated in Brazil since 1990 and is associated with severe disease and explosive outbreaks. Intending to shed light on the viral determinants for severe dengue pathogenesis, we sought to analyze the DENV-2 intrahost genetic diversity in 68 patient cases clinically classified as dengue fever (n = 31), dengue with warning signs (n = 19), and severe dengue (n = 18). Unlike previous DENV intrahost diversity studies whose approaches employed PCR, here we performed viral whole-genome deep sequencing from clinical samples with an amplicon-free approach, representing the real intrahost diversity scenario. Striking differences were detected in the viral population structure between the three clinical categories, which appear to be driven mainly by different infection times and selection pressures, rather than being linked with the clinical outcome itself. Diversity in the NS2B gene, however, showed to be constrained, irrespective of clinical outcome and infection time. Finally, 385 non-synonymous intrahost single-nucleotide variants located along the viral polyprotein, plus variants located in the untranslated regions, were consistently identified among the samples. Of them, 124 were exclusively or highly detected among cases with warning signs and among severe cases. However, there was no variant that by itself appeared to characterize the cases of greater severity, either due to its low intrahost frequency or the conservative effect on amino acid substitution. Although further studies are necessary to determine their real effect on viral proteins, this heightens the possibility of epistatic interactions. The present analysis represents an initial effort to correlate DENV-2 genetic diversity to its pathogenic potential and thus contribute to understanding the virus's dynamics within its human host.
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Vírus da Dengue/genética , Vírus da Dengue/isolamento & purificação , Dengue/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil , Criança , Pré-Escolar , Dengue/imunologia , Vírus da Dengue/classificação , Vírus da Dengue/fisiologia , Feminino , Variação Genética , Genoma Viral , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Filogenia , Sorogrupo , Proteínas não Estruturais Virais/genética , Adulto JovemRESUMO
Emerging and re-emerging viruses are a global health concern. Genome sequencing as an approach for monitoring circulating viruses is currently hampered by complex and expensive methods. Untargeted, metagenomic nanopore sequencing can provide genomic information to identify pathogens, prepare for or even prevent outbreaks. SMART (Switching Mechanism at the 5' end of RNA Template) is a popular approach for RNA-Seq but most current methods rely on oligo-dT priming to target polyadenylated mRNA molecules. We have developed two random primed SMART-Seq approaches, a sequencing agnostic approach 'SMART-9N' and a version compatible rapid adapters available from Oxford Nanopore Technologies 'Rapid SMART-9N'. The methods were developed using viral isolates, clinical samples, and compared to a gold-standard amplicon-based method. From a Zika virus isolate the SMART-9N approach recovered 10kb of the 10.8kb RNA genome in a single nanopore read. We also obtained full genome coverage at a high depth coverage using the Rapid SMART-9N, which takes only 10 minutes and costs up to 45% less than other methods. We found the limits of detection of these methods to be 6 focus forming units (FFU)/mL with 99.02% and 87.58% genome coverage for SMART-9N and Rapid SMART-9N respectively. Yellow fever virus plasma samples and SARS-CoV-2 nasopharyngeal samples previously confirmed by RT-qPCR with a broad range of Ct-values were selected for validation. Both methods produced greater genome coverage when compared to the multiplex PCR approach and we obtained the longest single read of this study (18.5 kb) with a SARS-CoV-2 clinical sample, 60% of the virus genome using the Rapid SMART-9N method. This work demonstrates that SMART-9N and Rapid SMART-9N are sensitive, low input, and long-read compatible alternatives for RNA virus detection and genome sequencing and Rapid SMART-9N improves the cost, time, and complexity of laboratory work.
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OBJECTIVES: This study was performed to determine whether Dengue virus (DENV) immunochromatographic tests can detect and differentiate nonstructural protein 1 (NS1) from each of the four DENV serotypes and do not cross-react with the Zika virus (ZIKV) NS1 protein. METHODS: We compared the specificity of six NS1-based DENV immunochromatographic tests (point of care) in the detection of NS1 proteins from each of the four DENV serotypes and ZIKV. The tests were performed with NS1 proteins produced in mammalian cells. Cross-reactivity was confirmed with a purified recombinant ZIKV NS1 protein and DENV+ or ZIKV+ human serum samples. RESULTS: Cross-reaction was observed in 2 out of the 6 evaluated tests using cell culture supernatants containing NS1 protein of each tested virus. Cross-reactivity with ZIKV was confirmed with purified recombinant ZIKV NS1 produced in Escherichia coli. Further analyses with serum samples collected from DENV+ or ZIKV+ patients confirmed the cross-reactivity with ZIKV protein in 2 tests. CONCLUSIONS: The detection of the NS1 protein is the basis for several commercially available serological DENV diagnostic tests. The present results emphasize the relevance of testing specificity of presently available NS1-based DENV serological tests and the need of adjustments of tests that cross-react with the ZIKV protein. Our results are particularly relevant for regions where both viruses are endemically found, as in the case of Brazil.
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Cromatografia de Afinidade/métodos , Vírus da Dengue/imunologia , Dengue/virologia , Proteínas não Estruturais Virais/imunologia , Zika virus/imunologia , Anticorpos Antivirais/sangue , Brasil , Reações Cruzadas , Vírus da Dengue/isolamento & purificação , Glicoproteínas/imunologia , Humanos , Sensibilidade e Especificidade , Especificidade da EspécieRESUMO
In Brazil, Dengue (DENV) and Zika (ZIKV) viruses are reported as being transmitted exclusively by Aedes aegypti in urban settings. This study established the vectors and viruses involved in an arbovirus outbreak that occurred in 2019 in a rural area of Espírito Santo state, Brazil. Mosquitoes collected were morphologically identified, sorted in samples, and submitted to molecular analysis for arboviruses detection. Phylogenetic reconstruction was performed for the viral sequence obtained. All 393 mosquitoes were identified as Aedes albopictus. DENV-1 genotype V was present in one sample and another sample was positive for ZIKV. The DENV-1 clustered with viruses that have circulated in previous years in large urban centers of different regions in Brazil. This is the first report of A. albopictus infected by DENV and ZIKV during an outbreak in a rural area in Brazil, indicating its involvement in arboviral transmission. The DENV-1 strain found in the A. albopictus was not new in Brazil, being involved previously in epidemics related to A. aegypti, suggesting the potential to A. albopictus in transmitting viruses already circulating in the Brazilian population. This finding also indicates the possibility of these viruses to disperse across urban and rural settings, imposing additional challenges for the control of the diseases.
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Aedes/virologia , Vírus da Dengue/isolamento & purificação , Dengue/transmissão , Mosquitos Vetores/virologia , Infecção por Zika virus/transmissão , Zika virus/isolamento & purificação , Animais , Brasil , Surtos de Doenças , Humanos , FilogeniaRESUMO
The present cohort study was set up with the aim of determining the incidence of dengue among children and adolescents, from 2 to 16 years of age, living in Araraquara, South-Eastern Brazil, a city classified as a mid-level endemicity location for dengue. Enrollment took place from September 2014 to March 2015. Baseline socio-demographic data were collected, and a blood sample from the participant was drawn, for dengue serology. Families were contacted weekly for fever surveillance. If the child developed fever, a nurse visited the household to collect a blood sample. PCR, NS1 and IgM were used for dengue diagnosis. Parents or legal guardians of participating children provided a written informed consent. 3,514 children and adolescents were enrolled in the cohort. Dengue baseline seroprevalence was 12.2% (95%CI: 11.1 - 13.3). The incidence density of symptomatic dengue was 8.94 per 100 person/years in the first year of follow-up, 0.58 in the second, and 0.19 in the fourth. No cases were confirmed in the third year. Incidence was associated with age, sex, baseline seroprevalence and with living in a house as opposed to an apartment. This study provides relevant information on the epidemiology of dengue in mid-level transmission settings that may be useful to policymakers in the evaluation of control strategies.
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Dengue/epidemiologia , Estudos Soroepidemiológicos , Adolescente , Adulto , Anticorpos Antivirais/sangue , Brasil/epidemiologia , Criança , Pré-Escolar , Estudos de Coortes , Dengue/virologia , Vírus da Dengue/genética , Feminino , Febre/epidemiologia , Humanos , Incidência , Lactente , MasculinoRESUMO
ABSTRACT We conducted a spatial case-control study nested in a dengue incidence cohort to explore the role of the spatial and socioeconomic factors in the proportion of symptomatic (cases) and inapparent primary dengue virus infections (controls). Cohort participants were children and adolescents (2 to 16 years of age) at the beginning of the follow-up. Case definitions were, for symptomatic cases, fever plus a positive lab result for acute dengue (NS1, RT-PCR, ELISA IgM/IgG), and for inapparent infection a positive result for dengue IgG (ELISA) in subjects without symptoms and with a previously negative result at baseline. The covariates included sociodemographic factors, residential location, and socioeconomic context variables of the census tracts of residence of cases and controls. We used principal component analysis to reduce the contextual covariates, with the component values assigned to each one based on their residences. The data were modeled in a Bayesian context, considering the spatial dependence. The final sample consisted of 692 children, 274 cases and 418 controls, from the first year of follow-up (2014-2015). Being male, older age, higher educational level of the head of the family and having a larger number of rooms in the household were associated with a greater chance of presenting dengue symptomatic infection at the individual level. The contextual covariates were not associated with the outcome. Inapparent dengue infection has extensive epidemiological consequences. Relying solely on notifications of symptomatic dengue infections underestimates the number of cases, preserves a silent source of the disease, potentially spreading the virus to unaffected areas.
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OBJECTIVES: This study was performed to determine whether neutralizing antibodies against yellow fever virus (YFV) generated by YFV vaccine could interfere in the specificity of dengue virus (DENV) and Zika virus (ZIKV) IgG ELISA tests. METHODS: Seventy-nine pairs of serum samples (pre- and post-vaccination), collected during the years 1997-1998 from children with no history of yellow fever disease who had been vaccinated against YFV, were tested. The seroconversion post-vaccination was evaluated through plaque reduction neutralization test (PRNT), and four different commercial ELISA kits were used for the detection of DENV and ZIKV IgG antibodies. RESULTS: A cross-reactivity rate of 3.9% with DENV IgG antibodies was found only with the Dengue Virus IgG Dx Select kit (Focus Diagnostics). CONCLUSIONS: As several countries have local transmission of multiple arboviruses, the absence of cross-reactivity or minimum cross-reactivity of YFV neutralizing antibodies with DENV and ZIKV antigens is a relevant finding, since the interpretation of sero-epidemiological investigations would be seriously impacted in many regions where YFV vaccination is mandatory.
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Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/sangue , Vírus da Dengue/imunologia , Vacina contra Febre Amarela/imunologia , Zika virus/imunologia , Antígenos Virais , Criança , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/sangue , Testes de Neutralização , Vírus da Febre Amarela/imunologiaRESUMO
Zika virus (ZIKV) clinical presentation and frequency/duration of shedding need further clarification. Symptomatic ZIKV-infected individuals identified in two hospitals in Sao Paulo State, Brazil, were investigated regarding clinical characteristics, shedding in body fluids, and serodynamics. Ninety-four of 235 symptomatic patients (Site A: 58%; Site B: 16%) had Real-Time PCR-confirmed ZIKV infection; fever, headache and gastrointestinal symptoms were less frequent, and rash was more frequent compared to ZIKV-negative patients. Real-Time PCR in serum had worse performance compared to plasma, while urine had the highest sensitivity. Shedding in genital fluids and saliva was rare. IgM positivity was the highest <14 days after the symptoms onset (86%), decreasing >28 days (24%); IgG positivity increased >14 days (96%) remaining positive in 94% of patients >28 days. ZIKV prevalence varied importantly in two neighboring cities during the same transmission season. Urine Real-Time PCR can improve diagnostic sensitivity; serum testing is less useful. Accurate serological tests are needed to improve diagnosis and surveillance.
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Secreções Corporais/virologia , Infecção por Zika virus/diagnóstico , Zika virus/isolamento & purificação , Adulto , Brasil/epidemiologia , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Gravidez , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Carga Viral , Infecção por Zika virus/epidemiologiaRESUMO
Dengue virus (DENV) and Zika virus (ZIKV) are members of the Flaviviridae and are predominantly transmitted via mosquito bites. Both viruses are responsible for a growing number of infections in tropical and subtropical regions. DENV infection can cause lethargy with severe morbidity and dengue shock syndrome leading to death in some cases. ZIKV is now linked with Guillain-Barré syndrome and fetal malformations including microcephaly and developmental disorders (congenital Zika syndrome). The protective and pathogenic roles played by the immune response in these infections is unknown. Mucosal-associated invariant T (MAIT) cells are a population of innate T cells with potent anti-bacterial activity. MAIT cells have also been postulated to play a role in the immune response to viral infections. In this study, we evaluated MAIT cell frequency, phenotype, and function in samples from subjects with acute and convalescent DENV infection. We found that in acute DENV infection, MAIT cells had elevated co-expression of the activation markers CD38 and HLA-DR and had a poor IFNγ response following bacterial stimulation. Furthermore, we found that MAIT cells can produce IFNγ in response to in vitro infection with ZIKV. This MAIT cell response was independent of MR1, but dependent on IL-12 and IL-18. Our results suggest that MAIT cells may play an important role in the immune response to Flavivirus infections.