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Enterovirus (EV) is commonly associated with central nervous system (CNS) syndromes. Recently, gastroenteric viruses, including rotavirus (RVA), human astrovirus (HAstV), and norovirus (NoV), have also been associated with CNS neurological disorders. The aim of the present study was to investigate the presence of EV, RVA, HAst, and NoV associated to CNS infections with undiagnosed etiology in Northwest region of São Paulo State, Brazil, and to conduct the molecular characterization of the positive samples detected. A total of 288 cerebrospinal fluid samples collected from July to December 2017 were tested for EV and NoV by quantitative real-time polymerase chain reaction (RT-qPCR), HAstV by conventional RT-PCR, and RVA by enzyme-linked immunosorbent assay. Positive-EV samples were inoculated in cells lines, amplified by RT-PCR and sequenced. RVA, NoV, and HAstV were not detected. EV infection was detected in 5.5% (16/288), and five samples successful genotyped: echovirus 3 (E3) (1/5), coxsackie virus A6 (CVA6) (1/5), and coxsackie virus B4 (CVB4) (3/5). Meningitis was the main syndrome observed (12/16; 75%). CVA6, CVB4, and E3 were identified associated with aseptic meningitis. Reports of CVA6 associated with aseptic meningitis are rare, E3 had not been previously reported in Brazil, and epidemiological data on CVB4 in the country is virtually unknown. The present investigation illustrates the circulation of diverse EV types in a small regional sample set and in a short period of time, highlighting the importance of an active EV surveillance system in CNS infections. Enhanced understanding of undiagnosed CNS infections will assist in public health and health care planning.

Measles is an acute and highly contagious but vaccine-preventable infectious disease. Despite years of being considered eliminated, decreased vaccination rates have produced virus reemergence in several countries, including Brazil. Measles can be controlled through immunization programs, through which aim to achieve 95% coverage with two doses of the vaccine. Measles can also be controlled if suspected cases can be properly identified in order to contain outbreaks. This cross-sectional study determined the prevalence of measles antibodies and their correlation with rubella antibodies (resulting from the combination vaccine used in Brazil’s public immunization program) in individuals aged higher 10 years old in São José do Rio Preto, São Paulo State, Brazil, participants of a prospective cohort of arbovirosis surveillance before virus reemergence in the country. Our findings presented that 32.9% of individuals aged 10–40 years old had not antibodies against measles; 39.3% of total individuals with documented evidence of measles vaccination did not have anti-measles IgG, though only 20.2% of individuals with documented evidence of rubella vaccination lacked anti-rubella IgG. Besides, the most of measles cases reported in the city, following the virus spreading in the country, occurred especially in groups defined by us as susceptible. Because the combination MMR vaccine is part of Brazil’s national vaccine schedule, the possible reasons for this relatively high rate of seronegativity need to be investigated further, once that it reflects outbreak risk.

Made available in DSpace on 2022-04-29T08:45:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Purpose: Studies show that around 80% of Zika virus (ZIKV) infections are asymptomatic. The present study tested urine samples from volunteers, unsuspected of arboviral infection, which attended an emergency care unit (ECU) in Mirassol, Brazil, from March 2018 to April 2019. Methods: The volunteers were divided into two groups. The first group was composed of outpatients who were not suspected to have an arbovirus infection. This first group was subdivided into two subgroups: outpatients with and without arbovirus-like symptoms. The second group consisted of companions of outpatients treated at the ECU. The second group was also subdivided into two subgroups: totally asymptomatic individuals and those who had arbovirus-like symptoms. RNA was extracted from urine samples, followed by RT-qPCR for ZIKV. Results: We found that 11% (79/697) of the samples tested positive for ZIKV-RNA. Among the ZIKV-RNA-positive individuals, 16.5% (13/79) were companions, of which 61.5% (8/13) were totally asymptomatic and 38.5% (5/13) reported symptoms that could be suggestive of arbovirus infection. In addition, 83.5% (66/79) of the ZIKV-RNA-positive individuals were outpatients without a clinical diagnosis of arbovirus. Of these undiagnosed ZIKV-RNA-positive outpatients, 47% (31/66) had no arbovirus-related symptoms. Conclusion: Our study shows the effectiveness of urine as a non-invasive sample to detect the incidence of ZIKV infection. We also highlight the importance of ZIKV molecular diagnosis to aid public health surveillance and prevention of congenital Zika syndrome and other ZIKV-associated diseases. Laboratório de Estudos Genômicos Universidade Estadual Paulista (Unesp) Instituto de Biociências Letras e Ciências Exatas (IBILCE), SP Department of Molecular Biology Faculdade de Medicina de São José do Rio Preto (FAMERP), SP Laboratório de Pesquisas em Virologia (LPV) Faculdade de Medicina de São José do Rio Preto (FAMERP), SP Department of Cardiology and Cardiovascular Surgery Faculdade de Medicina de São José do Rio Preto (FAMERP), SP Laboratório de Estudos Genômicos Universidade Estadual Paulista (Unesp) Instituto de Biociências Letras e Ciências Exatas (IBILCE), Rua Cristóvão Colombo, 2265, SP Laboratório de Estudos Genômicos Universidade Estadual Paulista (Unesp) Instituto de Biociências Letras e Ciências Exatas (IBILCE), SP Laboratório de Estudos Genômicos Universidade Estadual Paulista (Unesp) Instituto de Biociências Letras e Ciências Exatas (IBILCE), Rua Cristóvão Colombo, 2265, SP CAPES: 001 FAPESP: 2014/22198-0 CAPES: 2046/2016 CNPq: 440723/2016-7

Mayaro virus (MAYV) is a neglected mosquito-borne alphavirus that causes illness similar to Chikungunya (CHIKV), Dengue (DENV) and Zika virus (ZIKV). Currently, there is no specific treatment or vaccine against MAYV infection. To develop an efficient antiviral screening assay for MAYV, we constructed the infectious clones of MAYV strain BeAr 20290 and its eGFP reporter virus. The reporter virus exhibited high replication capacity indistinguishable with the wild type MAYV, and was genetically stable within at least five rounds of passages in BHK-21 cell. The expression of eGFP correlated well with the viral replication. Using the known inhibitor ribavirin, we confirmed that the MAYV-eGFP reporter virus could be used for antiviral screening to identify the specific inhibitors against MAYV. Using the MAYV-eGFP based antiviral assay, we found that the compound 6-Azauridine which had antiviral activity against CHIKV and SFV, showed a significant inhibitory effect on MAYV replication.

Background São José do Rio Preto is one of the cities of the state of São Paulo, Brazil, that is hyperendemic for dengue, with the presence of the four dengue serotypes. Objectives: to calculate dengue seroprevalence in a neighbourhood of São José do Rio Preto and identify if socioeconomic and demographic covariates are associated with dengue seropositivity. Methods A cohort study to evaluate dengue seroprevalence and incidence and associated factors on people aged 10 years or older, was assembled in Vila Toninho neighbourhood, São José do Rio Preto. The participant enrolment occurred from October 2015 to March 2016 (the first wave of the cohort study), when blood samples were collected for serological test (ELISA IgG anti-DENV) and questionnaires were administrated on socio-demographic variables. We evaluated the data collected in this first wave using a cross-sectional design. We considered seropositive the participants that were positive in the serological test (seronegative otherwise). We modelled the seroprevalence with a logistic regression in a geostatistical approach. The Bayesian inference was made using integrated nested Laplace approximations (INLA) coupled with the Stochastic Partial Differential Equation method (SPDE). Results We found 986 seropositive individuals for DENV in 1322 individuals surveyed in the study area in the first wave of the cohort study, corresponding to a seroprevalence of 74.6% (95%CI: 72.2–76.9). Between the population that said never had dengue fever, 68.4% (566/828) were dengue seropositive. Older people, non-white and living in a house (instead of in an apartment), were positively associated with dengue seropositivity. We adjusted for the other socioeconomic and demographic covariates, and accounted for residual spatial dependence between observations, which was found to present up to 800 m. Conclusions Only one in four people aged 10 years or older did not have contact with any of the serotypes of dengue virus in Vila Toninho neighbourhood in São José do Rio Preto. Age, race and type of house were associated with the occurrence of the disease. The use of INLA in a geostatistical approach in a Bayesian context allowed us to take into account the spatial dependence between the observations and identify the associated covariates to dengue seroprevalence. Electronic supplementary material The online version of this article (10.1186/s12879-019-4074-4) contains supplementary material, which is available to authorized users.

Background: The recent introduction of new arboviruses in the Americas, as Zika virus (ZIKV) and Chikungunya virus (CHIKV), increased the risk of outbreaks and arboviral co-infections. Herein, we report twelve cases of co-infection of ZIKV and different DENV serotypes in a city located in the northwest region of São Paulo State, Brazil, which is hyper-endemic to Dengue. Methods: Between January and November 2016, 1254 suspected cases of arboviral infection were available by our surveillance program in São José do Rio Preto. All suspected patients were examined and, when they were arboviral disease-suspectd, had sera separated and viral RNA analyzed by PCR/qPCR assays to determine the diagnosis of DENV 1-4, ZIKV, or CHIKV in the same samples. After the molecular results, twelve patients with ZIKV-DENV coinfection were identified and their clinical and laboratory characteristics were described. Results: The mean between symptoms onset and collected sample of 3 days. DENV-1 was identified in seven co-infected patients and DEN2 in other five. Two patients presented alarm signs of Dengue and no one was hospitalized. Conclusions: The constant presence of co-circulating arboviruses increases the chance of co-infection and demonstrates the importance of the differential diagnosis, especially during periods of arboviral outbreaks. The impact of this co-infection is known individual and collectively. Keywords: Arbovirus, Arboviruses, Co-infection, Dengue, Zika

Background The emergence of Zika virus (ZIKV) presents new challenges to both clinicians and public health authorities. Overlapping clinical features between the diseases caused by ZIKV, dengue (DENV) and chikungunya (CHIKV) and the lack of validated serological assays for ZIKV make accurate diagnosis difficult. Brazilian authorities largely rely on clinical and epidemiological data for the epidemiological and clinical classifications of most ZIKV cases. Objective : To report the laboratory and clinical profiles of patients diagnosed with Zika fever based only on clinical and epidemiological data. Study design We analyzed 433 suspected cases of ZIKV identified by the attending physician based on proposed clinical criteria. The samples were also screened for ZIKV, DENV and CHIKV using PCR. Results Of the 433 patients analyzed, 168 (38.8%) were laboratory-confirmed for arboviruses: 96 were positive for ZIKV, 67 were positive for DENV (56 for DENV-2, 9 for DENV-1, and 2 for DENV-4), four were positive for co-infection with ZIKV/DENV-2, and one was positive for CHIKV. The most common signs or symptoms in the patients with laboratory-confirmed ZIKV were rash (100%), arthralgia (77.1%), fever (74.0%), myalgia (74.0%) and non-purulent conjunctivitis (69.8%). In patients with laboratory-confirmed DENV infections, the most frequently observed symptoms were rash (100%), fever (79.1%), myalgia (74.6%), headache (73.1%) and arthralgia (70.1%). The measure of association between clinical manifestations and laboratory manifestations among patients with ZIKV and DENV detected a statistically significant difference only in abdominal pain (p = 0.04), leukopenia (p = 0.003), and thrombocytopenia (p = 0.01). Conclusion Our data suggests that clinical and epidemiological criteria alone are not a good tool for ZIKV and DENV differentiation, and that laboratory diagnosis should be mandatory.

Made available in DSpace on 2018-11-26T15:47:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01. Added 1 bitstream(s) on 2019-10-09T18:25:22Z : No. of bitstreams: 1 S1517-83822018000100144.pdf: 591481 bytes, checksum: 392b744e5bdcc30f75d329a1ae39d213 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Many countries in the Americas have detected local transmission of multiple arboviruses that cause febrile illnesses. Therefore, laboratory testing has become an important tool for confirming the etiology of these diseases. The present study aimed to compare the sensitivity and specificity of three different Zika virus detection assays. One hundred serum samples from patients presenting with acute febrile symptoms were tested using a previously reported TaqMan (R) RT-qPCR assay. We used a SYBR (R) Green RT-qPCR and a conventional PCR methodologies to compare the results. Of the samples that were determined to be negative by the TaqMan (R) RT-qPCR assay, 100% (Kappa = 0.670) were also found to be negative by SYBR (R) Green RT-qPCR based on Tm comparison; however, 14% (Kappa = 0.035) were found to be positive by conventional PCR followed by agarose gel electrophoresis. The differences between the ZIKV strains circulating worldwide and the low viremia period can compromise diagnostic accuracy and thereby the accuracy of outbreak data. Therefore, improved assays are required to improve the diagnosis and surveillance of arbovirus. (C) 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. Fac Med Sao Jose do Rio Preto FAMERP, Sao Jose Do Rio Preto, SP, Brazil Univ Estadual Paulista, Inst Biociencias Botucatu, Lab Virol, Botucatu, SP, Brazil Univ Nova Lisboa, GHTM, Lisbon, Portugal Prefeitura Sao Jose do Rio Preto, Dept Vigilancia Saude, Sao Jose Do Rio Preto, SP, Brazil Univ Estadual Paulista, Inst Biociencias Botucatu, Lab Virol, Botucatu, SP, Brazil FAPESP: 2013/21719-3

The Zika virus (ZIKV) epidemic in the Americas poses a public health emergency that requires a swift response. Accurate and reliable ZIKV diagnostic tests serve as an important tool for limiting the spread of ZIKV infections. The Aptima Zika virus assay (Hologic, Marlborough, MA) performed on the automated Panther system is a rapid and high-throughput method for detecting ZIKV RNA using transcription-mediated amplification (TMA) technology. We evaluated the performance characteristics of the Aptima Zika virus assay on clinical serum and urine specimens ( n = 124) from two different patient populations and samples spiked with ZIKV from three different lineages ( n = 10). Compared to the real-time reverse transcription-PCR (rRT-PCR) reference method, the Aptima ZIKV assay detected ZIKV RNA with a diagnostic accuracy of 94.8% (95% confidence interval [CI], 89.4 to 97.6), a sensitivity of 94.7% (95% CI, 73.5 to 99.9), and a specificity of 94.8% (95% CI, 88.9 to 97.8). Similar results were obtained regardless of whether a serum or urine source was used. The limits of detection of the assay at a 95% detection probability were 11.5 genome copy equivalents (GCE)/ml (95% fiducial limits, 7.9 to 20.2) in serum and 17.9 GCE/ml (95% fiducial limits, 13.1 to 27.5) in urine. The Aptima Zika virus assay results were highly reproducible (99%), and no cross-reactivity was seen during the testing of a panel of 95 specimens with potentially interfering substances, such as clinically relevant bacteria, fungi, and viruses, including other flaviviruses. The excellent performance characteristics and the convenience of a fully automated testing system make the Aptima ZIKV assay an attractive choice for clinical laboratories detecting ZIKV RNA from serum and urine.

Public health concerns exist surrounding the epidemic of the Zika virus (ZIKV) and the rapid growth of transplantation in developing countries, including endemic zones of active arbovirus transmission, as well as travel to such regions by potential organ donors and recipients. Few data exist regarding the clinical characteristics of ZIKV infection in immunocompromised hosts. Laboratory screening protocols for transplantation to differentiate ZIKV infections from other endemic viral diseases and for the detection of possible donor-derived infection have not been stated. The diagnosis of ZIKV infection remains a challenge, fueled by the lack of standardized commercially available diagnostic tests and validated reference diagnostic laboratories, as well as the limited duration of ZIKV viremia. In this small series, ZIKV infection in renal and liver recipients presented without rash, conjunctivitis, or neurological symptoms, and with abnormal graft function, thrombocytopenia, and bacterial superinfection. We report the first case series of ZIKV infection in solid organ recipients, with a description of clinical and laboratory features and therapeutic management.

Over 400 million people are estimated to be at risk of acquiring dengue virus (DENV). Despite efforts to mitigate the impact of DENV epidemics, the virus remains a public health problem in the Americas: more than one million DENV cases were reported in the continent between January and July 2019 DENV was first detected in Brazil in 1982, and Brazil has reported 88% (1,127,244 cases) of all DENV cases in the Americas during 2019 to date. São Paulo state in the southeast of Brazil has reported nearly half of all DENV infections in the country. Here we characterised the genetic diversity of DENV strains circulating in São Paulo state in 2019, at the epicentre of the ongoing DENV epidemic. Using portable nanopore sequencing we generated 20 new DENV genome sequences from viremic patients with suspected dengue infection residing in two of the most-affected municipalities, 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. The new outbreak strains were classified as DENV2 genotype III (American/Asian genotype). Notably, phylogenetic analysis indicated that the 2019 outbreak is the result of a novel DENV lineage that was recently introduced to Brazil from the Caribbean region. Our genetic analysis further indicates that the introduction and onwards spread of the outbreak lineage (named here DENV2-III BR-4) indicates a new DENV2 lineage replacement in Brazil. Dating phylogeographic analysis suggests that DENV2-III BR-4 was introduced to Brazil in or around early 2014, possibly from the Caribbean region. Our study describes the early detection of a newly introduced and rapidly-expanding DENV2 virus lineage in Brazil.Author SummaryDengue is the most important mosquito-borne viral disease of humans. The disease is caused by the dengue virus (DENV) that is classified within genus Flavivirus. DENV infections are caused by 4 serotypes (DENV 1-4) that are genetically related but antigenically distinct. Dengue infection results in a variety of symptoms that range from mild fever to dengue hemorrhagic fever and/or dengue shock syndrome (DHF/DSS). Clinical outcomes are associated with different types of infection, viral serotypes, genotypes, lineages, and host genetic factors. As a re-emerging infectious disease, DENV has become a serious threat to public health in the Americas, and particularly in Brazil, where it was introduced in the 1980s and became well established due to the country-wide re-infestation of the Aedes aegypti mosquito vector species. During the first six months of 2019, 1,282,183 DENV cases were reported in the Americas, with Brazil reporting a staggering 1,127,244 (88%) of all dengue cases in the continent. To date, no information exists on the genetic composition of the DENV lineage or lineages causing the current epidemic. Here we use portable sequencing to rapidly generate virus genome data from cases occurring in two different are severely-affected municipalities in São Paulo state, Brazil. We find that the 2019 dengue outbreak in Brazil is caused by a newly introduced DENV serotype 2 genotype III (Asian/American) that seems to be replacing previously-circulating DENV2 lineages. We discuss the potential implications of our results regarding the current outbreak in the context of previous outbreaks in the same region.

The disease caused by each of the four serotypes of dengue virus (DENV) have plagued humans since last century. Symptoms of dengue virus (DENV) infection range from asymptomatic to dengue fever (DF) to severe dengue disease (SDD). One third of the world's population lives in regions with active urban DENV transmission, and thousands of serologically naive travelers visit these areas annually, making a significant portion of the human population at risk of being infected. Even though lifelong immunity to the homotypic serotype is achieved after a primary DENV infection. Heterotypic DENV infections may be exacerbated by a pre-existing immune memory to the primary infection and can result in an increased probability of severe disease. Not only, age, comorbidities and presence of antibodies transferred passively from dengue-immune mother to infants are considered risk factors to dengue severe forms. Plasma leakage and multiple organ impairment are well documented in the literature, affecting liver, lung, brain, muscle, and kidney. However, unusual manifestation, severe or not, have been reported and may require medical attention. This review will summarize and discuss the increasing reports of unusual manifestations in the clinical course of dengue infection.

Arboviruses pose a serious threat to public health worldwide, overloading the healthcare system and causing economic losses. These viruses form a very diverse group, and in Brazil, arboviruses belonging to the families Flaviviridae and Togaviridae are predominant. Unfortunately, the number of arboviruses increases in proportion with factors such as deforestation, poor sanitation, climate changes, and introduction of new viruses like Chikungunya virus and Zika virus. In Brazil, dengue is endemic, along with the presence of other arboviruses. The situation is complicated by the scarcity of diagnostic infrastructure and the absence of approved vaccines for these diseases. Disease control, thus, relies solely on vector control. Therefore, enhanced clinical knowledge and improved general awareness about these arboviruses are indispensable to tackle diagnostic inadequacies.

Zika virus (ZIKV) infection acquired during pregnancy is associated with congenital microcephaly. We describe 2 cases of ZIKV infection in women in their 36th week of pregnancy whose fetuses had preserved head circumference at birth and findings of subependymal cysts and lenticulostriate vasculopathy in postnatal imaging. These represent the first signs of congenital brain injury acquired due to ZIKV in the third trimester.

During the 2016 Zika virus outbreak in Brazil, we detected Zika virus RNA in urine samples collected from Zika virus–positive pregnant women during different stages of pregnancy. Women had positive and negative intervals of viruria; 3 newborns had adverse outcomes. Further research is needed to clarify the relationship between viruria and outcomes for newborns.

Made available in DSpace on 2019-10-06T16:02:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 National Institutes of Health In Africa, Old World Primates are involved in the maintenance of sylvatic circulation of ZIKV. However, in Brazil, the hosts for the sylvatic cycle remain unknown. We hypothesized that free-living NHPs might play a role in urban/periurban ZIKV dynamics, thus we undertook an NHP ZIKV investigation in two cities in Brazil. We identified ZIKV-positive NHPs and sequences obtained were phylogenetically related to the American lineage of ZIKV. Additionally, we inoculated four C. penicillata with ZIKV and our results demonstrated that marmosets had a sustained viremia. The natural and experimental infection of NHPs with ZIKV, support the hypothesis that NHPs may be a vertebrate host in the maintainance of ZIKV transmission/circulation in urban tropical settings. Further studies are needed to understand the role they may play in maintaining the urban cycle of the ZIKV and how they may be a conduit in establishing an enzootic transmission cycle in tropical Latin America. São José do Rio Preto School of Medicine (FAMERP) Avenida Brigadeiro Faria Lima 5416 CEP: 15090-000, Vila São Pedro Laboratório de Vírus - Universidade Federal de Minas Gerais (UFMG) Avenida Antônio Carlos 6627 CEP: 31270-901 Pampulha Center for Drug Research and Development Instituto de Ciências Biológicas Universidade Federal de Minas Gerais (UFMG) Avenida Antônio Carlos 6627 CEP: 31270-901 Pampulha Instituto Adolfo Lutz (IAL) Avenida Dr. Arnaldo 351 − 7 Andar Sala 706 CEP: 01246-000 Pacaembú Laboratory of Wildlife Comparative Pathology Department of Pathology School of Veterinary Medicine and Animal Sciences University of São Paulo (LAPOCM-FMVZ-USP) Avenida Orlando Marques de Paiva 87 CEP: 05508-270 Department of Biology Institute of Biosciences Letters and Exact Sciences – São Paulo State University São José do Rio Preto – (IBILCE/UNESP), Rua Cristóvão Colombo, 2265, CEP: 15054-000 Epidemiological Surveillance Departament of São José do Rio Preto Avenida Romeu Strazzi 199 CEP: 15084-010 Vila Sinibaldi São Paulo State University (Unesp) Institute for Biotechnology Alameda das Tecomarias s/n CEP: 18607-440 Chácara Capão Bonito Department of Biochemistry and Molecular Biology University of Texas Medical Branch 301 University Blvd Centro de Controle de Zoonoses Belo Horizonte Council Rua Édna Quintel 173 CEP: 31270-705 São Bernardo Faceres Medical School Avenida Anísio Haddad 6751 CEP: 15090-305 Jardim Francisco Fernandes Department of Epidemiology School of Public Health of the University of São Paulo Avenida Dr. Arnaldo 715 CEP: 01246-904 Department of Pathology and Center of Biodefense and Emerging Infectious Diseases Center for Tropical Diseases Institute for Human Infections and Immunity University of Texas Medical Branch 301 University Blvd Department of Biology Institute of Biosciences Letters and Exact Sciences – São Paulo State University São José do Rio Preto – (IBILCE/UNESP), Rua Cristóvão Colombo, 2265, CEP: 15054-000 São Paulo State University (Unesp) Institute for Biotechnology Alameda das Tecomarias s/n CEP: 18607-440 Chácara Capão Bonito National Institutes of Health: R24AI120942

Madariaga virus (MADV), the new species designation for the South American isolates of eastern equine encephalitis virus (EEEV), is genetically divergent and substantially different in ecology and pathogenesis from North American EEEV strains. We isolated and characterized a MADV isolate obtained from a horse in Brazil. Our results support previous phylogenetic studies showing there are three genetically distinct MADV lineages. The MADV isolate from Paraíba State belongs to the South American lineage III and is closely related to Peruvian, Colombian and Venezuelan isolates.

St. Louis encephalitis virus (SLEV), a member of the family Flaviviridae, genus Flavivirus, is a causative agent of encephalitis in the Americas. In Brazil, sporadic cases of SLEV infection have been reported since 1953, but the first outbreak of SLEV in Brazil was identified only in 2007, concomitant with an outbreak of dengue virus (DENV) serotype 3. This finding, along with other reports, indicates that SLEV circulation in Brazil is largely unknown, and there may be epidemiological implications of the co-circulation of SLEV, DENV and other flaviviruses in Brazil. Here, we describe the first complete genome sequence of an SLEV strain isolated from a human patient in Brazil, strain BeH 355964. Phylogenetic analysis was performed to determine the genotype of BeH 355964 using the full-length genome and envelope (E) gene sequences separately. Both analyses showed that BeH 355964 could be classified as genotype V. Although the number of single gene sequences available is greater (such as for the E gene), the phylogenetic tree based on the complete genome sequence was better supported and provided further information about the virus.

Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of Mayaro fever, an acute febrile illness often presenting with arthralgic manifestations. The true incidence of MAYV infection is likely grossly underestimated because the symptomatic presentation is very similar to that of dengue fever and other acute febrile tropical diseases. We report the complete genome sequence of a MAYV isolate detected from an Acrelândia patient presenting with fever, chills, and sweating, but with no arthralgia. Results show that this isolate belongs to genotype D and is closely related to Bolivian strains. Our results suggest that the Acre/Mayaro strain is closely related to the progenitor of these Bolivian strains that were isolated between 2002 and 2006.

Made available in DSpace on 2018-11-26T17:41:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-15 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Butantan Institute Brazilian National Institutes for Science and Technology for Dengue Studies (INCT em Dengue) National Institutes of Health Background. The pathogenesis of severe dengue disease involves immune components as biomarkers. The mechanism by which some dengue virus (DENV)-infected individuals progress to severe disease is poorly understood. Most studies on the pathogenesis of severe dengue disease focus on the process of antibody-dependent enhancement (ADE) as a primary risk factor. With the circulation of Zika virus (ZIKV) in DENV-endemic areas, many people infected by ZIKV were likely exposed to DENV. The influence of such exposure on Zika disease outcomes remains unknown. Methods. We investigated whether patients previously exposed to DENV exhibited higher viremia when exposed to a subsequent, heterologous dengue or Zika infection than those patients not previously exposed to dengue. We measured viral loads and cytokine profile during patients' acute infections. Results. Neither dengue nor Zika viremia was higher in patients with prior DENV infection, although the power to detect such a difference was only adequate in the ZIKV analysis. Of the 10 cytokines measured, only 1 significant difference was detected: Levels of interleukin 1 beta (IL-1 beta) were lower in dengue-infected patients who had experienced a previous dengue infection than patients infected with dengue for the first time. However, power to detect differences between groups was low. In Zika-infected patients, levels of IL-1 beta showed a significant, positive correlation with viral load. Conclusions. No signs of ADE were observed in vivo in patients with acute ZIKV infection who had prior exposure to DENV. Sao Paulo State Univ, Sao Jose do Rio Preto Sch Med, Sao Jose Do Rio Preto, Brazil Sao Paulo State Univ, Butantan Inst, Sao Jose Do Rio Preto, Brazil Sao Paulo State Univ, Dept Biol, Inst Biosci Letters & Exact Sci, Sao Jose Do Rio Preto, Brazil New Mexico State Univ, Las Cruces, NM 88003 USA Univ Texas Med Branch, Galveston, TX 77555 USA Univ Sao Paulo, Sch Med, Sao Paulo, Brazil Sao Paulo State Univ, Sao Jose do Rio Preto Sch Med, Sao Jose Do Rio Preto, Brazil Sao Paulo State Univ, Butantan Inst, Sao Jose Do Rio Preto, Brazil Sao Paulo State Univ, Dept Biol, Inst Biosci Letters & Exact Sci, Sao Jose Do Rio Preto, Brazil FAPESP: 2013/21719-3 FAPESP: 2015/12295-0 National Institutes of Health: 1U01AI115577-01

Yellow fever virus (YFV) replication is highly dependent on host cell factors. YFV NS4B is reported to be involved in viral replication and immune evasion. Here interactions between NS4B and human proteins were determined using a GST pull-down assay and analyzed using 1-DE and LC-MS/MS. We present a total of 207 proteins confirmed using Scaffold 3 Software. Cyclophilin A (CypA), a protein that has been shown to be necessary for the positive regulation of flavivirus replication, was identified as a possible NS4B partner. 59 proteins were found to be significantly increased when compared with a negative control, and CypA exhibited the greatest difference, with a 22-fold change. Fisher's exact test was significant for 58 proteins, and the p value of CypA was the most significant (0.000000019). The Ingenuity Systems software identified 16 pathways, and this analysis indicated sirolimus, an mTOR pathway inhibitor, as a potential inhibitor of CypA. Immunofluorescence and viral plaque assays showed a significant reduction in YFV replication using sirolimus and cyclosporine A (CsA) as inhibitors. Furthermore, YFV replication was strongly inhibited in cells treated with both inhibitors using reporter BHK-21-rep-YFV17D-LucNeoIres cells. Taken together, these data suggest that CypA-NS4B interaction regulates YFV replication. Finally, we present the first evidence that YFV inhibition may depend on NS4B-CypA interaction.

The recent Zika virus (ZIKV) outbreak demonstrates that cost-effective clinical diagnostics are urgently needed to detect and distinguish viral infections to improve patient care. Unlike dengue virus (DENV), ZIKV infections during pregnancy correlate with severe birth defects, including microcephaly and neurological disorders. Because ZIKV and DENV are related flaviviruses, their homologous proteins and nucleic acids can cause cross-reactions and false-positive results in molecular, antigenic, and serologic diagnostics. We report the characterization of monoclonal antibody pairs that have been translated into rapid immunochromatography tests to specifically detect the viral nonstructural 1 (NS1) protein antigen and distinguish the four DENV serotypes (DENV1-4) and ZIKV without cross-reaction. To complement visual test analysis and remove user subjectivity in reading test results, we used image processing and data analysis for data capture and test result quantification. Using a 30-μl serum sample, the sensitivity and specificity values of the DENV1-4 tests and the pan-DENV test, which detects all four dengue serotypes, ranged from 0.76 to 1.00. Sensitivity/specificity for the ZIKV rapid test was 0.81/0.86, respectively, using a 150-μl serum input. Serum ZIKV NS1 protein concentrations were about 10-fold lower than corresponding DENV NS1 concentrations in infected patients; moreover, ZIKV NS1 protein was not detected in polymerase chain reaction-positive patient urine samples. Our rapid immunochromatography approach and reagents have immediate application in differential clinical diagnosis of acute ZIKV and DENV cases, and the platform can be applied toward developing rapid antigen diagnostics for emerging viruses.

Made available in DSpace on 2018-12-11T17:03:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-08-01 Background: The Zika virus (ZIKV) is an emerging arthropod-borne virus related to the dengue virus (DENV), and shows a similar clinical profile as other arboviral diseases, such as dengue and chikungunya virus (CHIKV). Historically, ZIKV has been associated with sporadic cases of human infection, but is now responsible for outbreaks worldwide. In Brazil, cases have been reported since 2015, with some cases causing severe disease. Objective: To identify clinical symptoms of Zika in patients in Dengue suspected patients. Study design: Description of a series of cases, wherein we analyzed 100 clinical samples collected from patients who exhibited acute febrile disease for ≤5 days, from January to February 2016. Results: In this study, we report 13 cases of ZIKV infection in adults presenting dengue-like symptoms in a DENV endemic area. All patients presented with fever, with myalgia being the second most frequently observed symptom. Two patients had rashes, but none of them had conjunctivitis. Other less frequent manifestations included headache, arthralgia, diarrhea, and nausea. Conclusion: The co-circulation of ZIKV and DENV is a serious public health concern, since it represents both a clinical and diagnostic challenge in endemic areas, as well as in the field of travel medicine. São José do Rio Preto School of Medicine (FAMERP) Avenida Brigadeiro Faria Lima, 5416. Vila São Pedro. Global Health and Tropical Medicine (GHTM) Universidade Nova de Lisboa, Rua da Junqueira No 100 São José do Rio Preto Regional School of Medicine Foundation (FUNFARME) Avenida Brigadeiro Faria Lima, 5416. Vila São Pedro. Department of Biology Institute of Biosciences Letters and Exact Sciences - São Paulo State University São José do Rio Preto - (IBILCE/UNESP), Rua Cristóvão Colombo, 2265 Department of Biology Institute of Biosciences Letters and Exact Sciences - São Paulo State University São José do Rio Preto - (IBILCE/UNESP), Rua Cristóvão Colombo, 2265

Dengue fever is the most common arbovirus disease, and presents with a large spectrum of clinical manifestations ranging from asymptomatic disease through to the development of dengue hemorrhagic fever. These extreme cases can lead to dengue shock syndrome, and sometimes death. Spinal cord involvement in dengue virus (DENV) infections is rare. Here, we report a case in which the patient developed acute transverse myelitis (TM) without paraparesis following a DENV infection. This case highlights the importance of physicians' awareness of the possible link between DENV and TM in endemic areas.

Arboviruses are frequently associated with outbreaks in humans and represent a serious public health problem. Among the Brazilian arboviruses, Mayaro virus, Dengue virus (DENV), Yellow Fever virus, Rocio virus, Saint Louis Encephalitis virus (SLEV), and Oropouche virus are responsible for most of human cases. All these arboviruses usually produce undistinguishable acute febrile illness, especially in the acute phase of infection. In this study we investigated the presence of arboviruses in sera of 519 patients presenting acute febrile illness, during a dengue outbreak in São José do Rio Preto City (São Paulo, Brazil). A multiplex-nested RT-polymerase chain reaction assay was applied to detect and identify the main Brazilian arboviruses (Flavivirus, Alphavirus, and Orthobunyavirus genera). The molecular analysis showed that 365 samples were positive to DENV-3, 5 to DENV-2, and 8 to SLEV. Among the positive samples, one coinfection was detected between DENV-2 and DENV-3. The phylogenetic analysis of the SLEV envelope gene indicated that the virus circulating in city is related to lineage V strains. These results indicated that during that large DENV-3 outbreak in 2006, different arboviruses cocirculated causing human disease. Thus, it is necessary to have an efficient surveillance system to control the dissemination of these arboviruses in the population.

To the Editor: Oropouche virus (OROV), a member of the Bunyaviridae family, Orthobunyavirus genus, Simbu serogroup, is transmitted to humans in urban areas by the biting midge Culicoides paraensis and causes epidemic acute febrile disease (1). Since its first isolation in Trinidad in 1955 (2), OROV has been associated with large outbreaks in South and Central America; half a million cases have been described during the past 45 years (1). The tripartite genome of OROV comprises single-strand, negative-sense large (L), medium (M), and small (S) RNAs that encode RNA polymerase, glycoproteins, and nucleocapsid, respectively. Studies have indicated the existence of 3 genotypes of OROV circulating in Brazil: genotypes I and II in the Amazon Basin and genotype III in the Southeast Region (3–5). OROV causes explosive urban epidemics. Serologic evidence of exposure to OROV in populations not affected by known outbreaks suggests that the virus circulates endemically (1). However, no sporadic infections have been reported. Here we report a sporadic OROV infection detected by clinical and laboratory surveillance of acute febrile illnesses in Acre, a state in the western Amazon region of Brazil. From March 2004 through October 2006, we prospectively investigated 69 febrile episodes in persons 6–60 years of age (mean, 28.1 years) living in the town of Acrelândia (10°13′W, 67°00′S) and surrounding rural areas (25.7% and 74.3% of the sample, respectively). Serum samples for reverse transcription–PCR (RT-PCR) were stored in liquid nitrogen in the field and shipped on dry ice to the laboratory in Sao Jose do Rio Preto, 3,500 km southeast of Acre. Because malaria and several arboviruses are locally endemic (6), all patients were screened for malarial parasites by thick-smear microscopy and for flaviviruses and alphaviruses by multiplex-nested RT-PCR (7). The samples negative for both malaria and other arboviruses were further tested for OROV with primers targeting the S segment of the OROV genome in a seminested RT-PCR strategy (R.V.M. Bronzoni et al., unpub. data; primers and protocol available from the authors by request). The sample also was isolated in Vero cells, and the RT-PCR described by Moreli et al. (8) was used for confirmation. We sequenced amplicons by using the same primers used for RT-heminested amplification and by using BigDye Terminators version 3.1 (ABI, Foster City, CA, USA) in ABI377 automated sequencer. Sequences were edited by DSGene 2.0 (Accelrys, San Diego, CA, USA) and deposited in GenBank (accession no. {"type":"entrez-nucleotide","attrs":{"text":"EU561644","term_id":"189212452","term_text":"EU561644"}}EU561644). One (1.4%) of 69 samples tested for OROV by heminested PCR was positive. This sample (BR/2004/ACRE27) was collected from a male patient from a rural area in April 2004. Precautions were followed to avoid contamination; positive and negative controls were used in all reactions; and the procedure was reproduced several times. The patient had ill-defined, mild flu-like symptoms; low-grade fever; and nasal discharge but reported no headache or other major symptoms. He recovered without complication. We built a phylogenetic tree on the basis of the 522 nucleotide sequences (27–200 aa) of nucleocapsid protein gene of OROV sample BR/2004/ACRE27 and other GenBank sequences from different OROV genotypes. We used sequences from Aino, Akabane, and Tinaroo viruses as the outgroup. A phylogenetic analysis was performed by the neighbor-joining method by using the Kimura 2-parameter nucleotide substitution model (9). The tree showed 3 main clades, corresponding to genotypes I, II, and III, and BR/2004/ACRE27 grouped within genotype I strains (Figure). Both genotypes I and II have been described in OROV outbreaks in Acre; genotype I, however, is found mostly in Para in the eastern part of the Brazilian Amazon region. Figure Phylogenetic tree of Oropouche virus strains; boldface shows the sample from the patient in this study. Phylogenetic tree was constructed from partial nucleocapsid gene sequence (522 nt, 27–200 aa) by neighbor-joining method implemented in MEGA ... A baseline serologic survey in rural Acrelândia during March and April 2004 detected antibodies to OROV in 6 (1.7%) of 357 persons 5–90 years of age who were examined by microplaque hemagglutination inhibition (10). Because none of these persons had been exposed to known OROV outbreaks in Acre or elsewhere, these findings further suggest the sporadic circulation of OROV in the area. We describe a sporadic infection of OROV infection in the Amazon region of Brazil in a mildly symptomatic patient. The nucleocapsid gene of the isolate has been sequenced, placing it in the genotype I group, the most commonly found in the Amazon Basin. These data suggest that OROV circulation may be sporadic and clinically silent and, when not associated with outbreaks, most likely neglected by local physicians.

Made available in DSpace on 2013-08-12T18:18:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-09-01 Made available in DSpace on 2013-09-30T18:41:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T14:01:07Z No. of bitstreams: 1 S0037-86822010000500007.pdf: 768765 bytes, checksum: 540d93f9d2064a901bd1f0948f4fbbe8 (MD5) Made available in DSpace on 2014-05-20T14:01:08Z (GMT). No. of bitstreams: 1 S0037-86822010000500007.pdf: 768765 bytes, checksum: 540d93f9d2064a901bd1f0948f4fbbe8 (MD5) Previous issue date: 2010-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Introduction: The Amazon region has extensive forested areas and natural ecosystems, providing favorable conditions for the existence of innumerous arboviruses. Over 200 arboviruses have been isolated in Brazil and about 40 are associated with human disease. Four out of 40 are considered to be of public health importance in Brazil: Dengue viruses (1-4), Oropouche, Mayaro and Yellow Fever. Along with these viruses, about 98% of the malaria cases are restricted to the Legal Amazon region. Methods: This study aimed to investigate the presence of arboviruses in 111 clinical serum samples from patients living in Novo Repartimento (Para), Placido de Castro (Acre), Porto Velho (Rondonia) and Oiapoque (Amapa). The viral RNA was extracted and RT-PCR was performed followed by a Multiplex-Nested-PCR, using Flavivirus, Alphavirus and Orthobunyavirus generic and species-specific primers. Results: Dengue virus serotype 2 was detected in two patients living in Novo Repartimento (Para) that also presented active Plasmodium vivax infection. Conclusions: Despite scant data, this situation is likely to occur more frequently than detected in the Amazon region. Finally, it is important to remember that both diseases have similar clinical findings, thus the diagnosis could be made concomitantly for dengue and malaria in patients living or returning from areas where both diseases are endemic or during dengue outbreaks. Sao Jose Rio do Preto Med Sch, Virol Lab, Sao Jose do Rio Preto, SP, Brazil Sao Jose Rio do Preto Med Sch, Dept Dermatol & Infect Dis, Sao Jose do Rio Preto, SP, Brazil São Paulo State Univ, Grad Program Microbiol, Sao Jose do Rio Preto, SP, Brazil São Paulo State Univ, Grad Program Microbiol, Sao Jose do Rio Preto, SP, Brazil FAPESP: 2008/03828-1 FAPESP: 2002/0946-1 CNPq: 302353/03-8

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:50:47Z No. of bitstreams: 1 S0100-736X2009000200015.pdf: 136539 bytes, checksum: b497e8a6e1b6134564b825c266e6298d (MD5) Made available in DSpace on 2013-08-22T18:50:47Z (GMT). No. of bitstreams: 1 S0100-736X2009000200015.pdf: 136539 bytes, checksum: b497e8a6e1b6134564b825c266e6298d (MD5) Previous issue date: 2009-02-01 Made available in DSpace on 2013-09-30T17:56:47Z (GMT). No. of bitstreams: 2 S0100-736X2009000200015.pdf: 136539 bytes, checksum: b497e8a6e1b6134564b825c266e6298d (MD5) S0100-736X2009000200015.pdf.txt: 34079 bytes, checksum: 061978ed7ca82bf1f7aca9c8832c637a (MD5) Previous issue date: 2009-02-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:16:13Z No. of bitstreams: 2 S0100-736X2009000200015.pdf: 136539 bytes, checksum: b497e8a6e1b6134564b825c266e6298d (MD5) S0100-736X2009000200015.pdf.txt: 34079 bytes, checksum: 061978ed7ca82bf1f7aca9c8832c637a (MD5) Made available in DSpace on 2014-05-20T13:16:13Z (GMT). No. of bitstreams: 2 S0100-736X2009000200015.pdf: 136539 bytes, checksum: b497e8a6e1b6134564b825c266e6298d (MD5) S0100-736X2009000200015.pdf.txt: 34079 bytes, checksum: 061978ed7ca82bf1f7aca9c8832c637a (MD5) Previous issue date: 2009-02-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O serpin maspin, um supressor tumoral no câncer de mama foi descrito como inibidor de migração celular e indutor de adesão celular entre a membrana basal e a matriz extracelular resultando na inibição da metástase tumoral. Por outro lado, a alta expressão do maspin está relacionada com um mau prognóstico em outros tipos de câncer. Pouco se sabe sobre a expressão, regulação e função do maspin nos tumores mamários caninos. Neste estudo, foi demonstrada uma perda da expressão de maspin nas células mamárias malignas de cães quando comparadas com um pool de tecido mamário normal de cães, analisado por PCR quantitativa em tempo real. Houve uma expressão fraca maspin em preparações de tumores mamários malignos observadas por imuno-histoquímica. Também foi verificado que a expressão nuclear do maspin em tumores mamários caninos está relacionada a um bom prognóstico. Assim, o maspin pode ser utilizado como um marcador prognóstico nas neoplasias mamárias em cães. The serpin maspin, a tumor suppressor in breast cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumor metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other types of cancer. Little is known about expression, regulation and function of maspin in canine mammary tumors. It was demonstrated in this study, a loss of maspin expression in malignant canine mammary cells compared with a pool of normal canine mammary tissue, analyzed by quantitative real-time PCR; weak maspin expression in malignant canine mammary tumors were observed by immunohistochemistry. It was also demonstrated that a correlation with nuclear maspin expression and a good prognosis. It is suggested that maspin could be used as a prognostic marker in canine mammary neoplasia. Faculdade de Medicina de São José do Rio Preto (FAMERP) Centro Regional de Bioterismo FAMERP Unidade de Pesquisa em Genética e Biologia Molecular Universidade Estadual Paulista Laboratório de Patologia Animal FAMERP Laboratório de Bioinformática FAMERP Laboratório de Pesquisa em Virologia Laboratório de Patologia e Biologia Molecular Veterinária Universidade Estadual Paulista Laboratório de Patologia Animal FAPESP: 05/51936-0

A apoptose e a proliferação celular possuem uma participação importante na tumorigênese, determinando o crescimento tumoral e consequentemente sua agressividade. O presente estudo teve como objetivo avaliar a ocorrência da apoptose associada a proliferação celular em neoplasias mamárias em cadelas e a elas a evolução clínica do paciente. Setenta animais foram submetidos à exérese cirúrgica do tumor, sendo este submetido ao diagnóstico histopatológico e marcação imuno-histoquímica para caspase-3 e Ki-67. Estes marcadores de apoptose e proliferação celular demonstraram grande expressão nas neoplasias malignas, principalmente nos carcinomas, considerado o mais maligno dos tumores. Estes resultados corroboram os dados da literatura e contribuem para um prognóstico tumoral criterioso que complementa a classificação tumoral existente proporcionando uma melhor e maior sobrevida devido a uma adequação do procedimento terapêutico de cada paciente.

Faculdade de Medicina de Sao Jose do Rio Preto (FAMERP) FAMERP, Lab Pesquisa Virol, Sao Jose do Rio Preto, Brazil

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:29:47Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:43:15Z : No. of bitstreams: 1 2-s2.0-84879126531.pdf: 2188616 bytes, checksum: 923e564ce1a446c1afc928f0fc430c35 (MD5) Made available in DSpace on 2014-05-27T11:29:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-25 Background: Yellow fever virus (YFV) belongs to the Flavivirus genus and causes an important disease. An alarming resurgence of viral circulation and the expansion of YFV-endemic zones have been detected in Africa and South America in recent years. NS5 is a viral protein that contains methyltransferase and RNA-dependent RNA polymerase (RdRp) domains, which are essential for viral replication, and the interactions between NS5 and cellular proteins have been studied to better understand viral replication. The aim of this study was to characterize the interaction of the NS5 protein with eukaryotic translation initiation factor 3 subunit L (eIF3L) and to evaluate the role of eIF3L in yellow fever replication. Methods. To identify interactions of YFV NS5 with cellular proteins, we performed a two-hybrid screen using the YFV NS5 RdRp domain as bait with a human cDNA library, and RNApol deletion mutants were generated and analyzed using the two-hybrid system for mapping the interactions. The RNApol region involved was segmented into three fragments and analyzed using an eIF3L-expressing yeast strain. To map the NS5 residues that are critical for the interactions, we performed site-direct mutagenesis in segment 3 of the interaction domain (ID) and confirmed the interaction using in vitro assays and in vivo coimmunoprecipitation. The significance of eIF3L for YFV replication was investigated using eIF3L overexpression and RNA interference. Results: In this work, we describe and characterize the interaction of NS5 with the translation factor eIF3L. The interaction between NS5 and eIF3L was confirmed using in vitro binding and in vivo coimmunoprecipitation assays. This interaction occurs at a region (the interaction domain of the RNApol domain) that is conserved in several flaviviruses and that is, therefore, likely to be relevant to the genus. eIF3L overexpression and plaque reduction assays showed a slight effect on YFV replication, indicating that the interaction of eIF3L with YFV NS5 may play a role in YFV replication. Conclusions: Although the precise function of eIF3L on interactions with viral proteins is not entirely understood, these results indicate an interaction of eIF3L with YF NS5 and that eIF3L overexpression facilitates translation, which has potential implications for virus replication. © 2013 Morais et al.; licensee BioMed Central Ltd. Laboratório de Pesquisas em Virologia Departamento de Doenças Dermatológicas, Infecciosas e Parasitárias Faculdade de Medicina de São José Do Rio Preto-FAMERP, Av. Brigadeiro Faria Lima 5416, São José do Rio Preto, SP 15090-000 Programa de Pós-graduação em Microbiologia Departamento de Biologia Campus São José Do Rio Preto-IBILCE/UNESP, 15054-000, São José do Rio Preto, SP Departamento de Virologia e Terapia Experimental Centro de Pesquisas Aggeu Magalhães CPqAM/FIOCRUZ, Av. Professor Moraes Rego s/n, Recife, PE 50670-420 Departamento de Ciências Biológicas Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista, Araraquara 14801-902 Center for Tropical Diseases University of Texas Medical Branch, Galveston, TX Programa de Pós-graduação em Microbiologia Departamento de Biologia Campus São José Do Rio Preto-IBILCE/UNESP, 15054-000, São José do Rio Preto, SP Departamento de Ciências Biológicas Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista, Araraquara 14801-902