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Background Acute respiratory infections caused by viruses are among the leading causes of morbidity and mortality. The inflammatory response that follows viral infection is important for the control of virus proliferation. However, if overwhelming, may be associated with complicated outcomes. Objectives We assessed the clinical characteristics of patients with severe acute respiratory illness (SARI) evolving to acute respiratory distress syndrome (ARDS) and the factors related to death. Study design. Prospective study in 273 adult patients with SARI performed in a university-affiliated 800-bed hospital serving an area of epidemiologic vigilance of 102 municipalities and more than 2 million inhabitants. Influenza A (H1N1) 2009 (A/H1N1), influenza A H3N2, and influenza B were tested in all patients by RT-PCR. Results The overall hospital mortality rate was 17.6%. A total of 30.4% of patients tested positive for influenza A/H1N1. Patients with SARI that evolved to ARDS took significantly longer to take the first dose of oseltamivir (6.0 vs 1.0 days, p=0.002). Patients with H1N1 positive tests had almost 3 times higher probability of death, despite having significantly less comorbidities (p=0.027). The influenza A/H1N1 pdm09 vaccine reduced the odds of death by 78%. Nonsurvivors had a more intense inflammatory response than did survivors at 48 h (C-reactive protein: 31.0 ± 17.5 vs. 14.6 ± 8.9 mg/dl, p=0.001) as well as a more positive fluid balance. Conclusions Hospital mortality associated with influenza H1N1-associated SARI and ARDS continued to be high years after the 2009 pandemic in a population with low vaccine coverage. Antiviral treatment started more than two days after onset of symptoms was more frequently associated with ARDS and death and, having had vaccine against influenza A (H1N1) was a factor independently related to survival.

Background The respiratory syncytial virus (RSV) is recognized as an important cause of respiratory tract infections. Immunocompromised patients, healthcare workers (HCWs) and children contacts are at increased risk of acquiring the infection. However, the impact of asymptomatic infection in transmission has not been well studied. Objectives: this study evaluated the frequency and viral load (VL) of RSV in nasal swab samples of individuals with different risk factors for acquiring infection in a university hospital in Sao Paulo, Brazil. Methods We included 196 symptomatic children and their 192 asymptomatic caregivers, 70 symptomatic and 95 asymptomatic HCWs, 43 samples from symptomatic HIV‐positive outpatients, and 100 samples of asymptomatic HIV patients in the period of 2009‐2013. Results RSV infection was detected in 10.1% (70/696) of samples, 4.4% (17/387) of asymptomatic patients, and 17.1% (53/309) from symptomatic patients. (P < .0001). The VL of symptomatic patients (4.7 log copies/mL) was significantly higher compared to asymptomatic patients (2.3 log copies/mL). RSV detection among asymptomatic caregivers (6.8%; 13/192) was significantly higher compared to other asymptomatic adults, HIV and HCWs (2.0%; 4/195; P = .0252). A close contact with an infected child at home was an important risk to RSV acquisition [OR 22.6 (95% CI 4.8‐106.7)]. Children who possibly transmitted the virus to their asymptomatic contacts had significantly higher viral load than children who probably did not transmit (P < .0001). Conclusions According to our results, it is important to know if people circulating inside the hospital have close contact with acute respiratory infected children.

Rotavirus is the main global cause of severe childhood diarrhoea among children. In 2006, Rotarix® (G1P[8]) was introduced into Brazil’s National Immunization Program. The vaccine coverage rate was 84.4% in 2009. Evidences of increasing G2P[4] after 2006 opened up the discussion about the vaccine effectiveness to non-G1 strains. The aim of this study was to identify the circulating rotavirus genotypes in two Brazilian regions during 2009. A total of 223 positive samples by immunochromatography and latex agglutination assay from the Northeast (Bahia/Pernambuco States) and Southeast (São Paulo/Rio de Janeiro States) regions were included in the study. The samples were submitted to genotyping by nested-PCR according to VP7(G) and VP4(P) and 175 samples (78.5%) were able to be characterized. Considering the characterization of VP7, the G-types detected were G1, G2, and G4 in the Northeast, and G2, G3, G5, and G9 in the Southeast. Considering the characterization of VP4, the P-types detected were P[4], P[8], and P[6]/P[9] in the Northeast and the Southeast. The most frequent mixed types found were G2P[4]/G2P[NT](81.4%), G2P[6](5.2%), G1P[6](5.2%) in the Northeast, and G2P[4]/G2P[NT](78.8%), G2P[6](8.2%), G9P[8](4.7%) in the Southeast. Among immunized individuals whose age ranged from 0-4 years, the G2P[4]/G2P[NT] genotype was identified in 91,0% of cases, and among non-immunized individuals of the same age, the G2P[4]/G2P[NT] genotype was identified in 85.7% of the cases. In accordance with the high level of vaccine coverage, the data suggest that the circulation of G2P[4] in these regions had a considerable increase after the introduction of Rotarix®.

The dynamics of dengue virus (DENV) circulation depends on serotype, genotype and lineage replacement and turnover. In São José do Rio Preto, Brazil, we observed that the L6 lineage of DENV-1 (genotype V) remained the dominant circulating lineage even after the introduction of the L1 lineage. We investigated viral fitness and immunogenicity of the L1 and L6 lineages and which factors interfered with the dynamics of DENV epidemics. The results showed a more efficient replicative fitness of L1 over L6 in mosquitoes and in human and non-human primate cell lines. Infections by the L6 lineage were associated with reduced antigenicity, weak B and T cell stimulation and weak host immune system interactions, which were associated with higher viremia. Our data, therefore, demonstrate that reduced viral immunogenicity and consequent greater viremia determined the increased epidemiological fitness of DENV-1 L6 lineage in São José do Rio Preto., Author summary Since 2008 L6 is the endemic lineage that has circulated in SJRP. In 2010, the L1 lineage was first identified in the city. For a period, both lineages co-circulated, and then in 2013, L1 began to diminish until it was no longer detected in the population. Differences in replicative fitness are usually the main factor for clade replacement (CR) in DENV epidemics. However, despite the better viral fitness of the emerging lineage, the absence of CR could not be explained by these differences alone. Here, we combine epidemiological, phylogenetic, molecular and immunological analyses to provide a more precise understanding of the role of fitness in lineage dynamics with the persistence of L6 even after the introduction of L1 without CR. Differences in immune responses elicited by DENV-1 L1 and L6 lineages (genotype V), but not viral fitness in mosquito or human cells, explain the dynamics of circulating DENV in a city of Brazil.

INTRODUCTION: Human adenoviruses (HAdV) play an important role in the etiology of severe acute lower respiratory infection, especially in immunocompromised individuals. The aim of the present study was detect the HAdV through different methods: direct fluorescence assay (DFA) and nested-polymerase chain reaction (PCR-nested) from patients with acute respiratory infection (ARI) up to 7 days of symptoms onset. METHODS: Samples (n=643) were collected from different risk groups during from 2001 to 2010: 139 adults attended in an Emergency Room Patients (ERP); 205 health care workers (HCW); 69 from Renal Transplant Outpatients (RTO); 230 patients in hematopoietic stem cell transplantation (HSCT) program. RESULTS: Among all patients (n=643) adenovirus was detected on 13.2% by DFA and/or PCR: 6/139 (4.3%) adults from ERP, 7/205 (3.4%) from HCW samples, 4/69 (5.8%) from RTO and 68/230 (29.5%) from HSCT patients. Nested PCR showed higher detection (10%) compared to DFA test (3.8%) (p < 0.001). HSCT patients presented significantly higher prevalence of HAdV infection. CONCLUSIONS: Adenovirus detection through nested-PCR assay was higher. However the inclusion of molecular method in laboratorial routine diagnostic should be evaluated considering the reality of each specific health service. Universidade Federal de São Paulo (UNIFESP) Departamento de Medicina Disciplina de Infectologia Universidade do Centro-Oeste Departamento de farmacologia Clínica UNIFESP, Depto. de Medicina Disciplina de Infectologia SciELO

INTRODUÇÃO E OBJETIVOS: O bocavírus humano (HBoV) tem sido descrito desde 2005 como agente etiológico de infecções respiratórias virais. O presente estudo tem como objetivo investigar a etiologia da infecção respiratória pelo HBoV em pacientes adultos e crianças de diferentes grupos de risco para complicação de infecções respiratórias agudas desde 2001 até 2008 em um hospital terciário no Brasil. PACIENTES E MÉTODOS: O HBoV foi investigado, através de reação em cadeia da polimerase, em 598 amostras respiratórias coletadas de pacientes hospitalizados e não hospitalizados. RESULTADOS: Das 598 amostras testadas o HBoV foi detectado em 2,44% (8/328) das crianças, incluindo cinco crianças portadoras de cardiopatia congênita, e 0,4% (1/270) dos adultos em programa de transplante de células tronco hematopoiéticas. CONCLUSÃO: Os dados do presente estudo sugerem baixa freqüência de detecção do HBoV entre pacientes de risco, e destaca a necessidade de novos estudos para um melhor entendimento do verdadeiro papel desse agente em infecções respiratórias agudas em pacientes sintomáticos. BACKGROUND AND OBJECTIVES: Human Bocavirus (HBoV) has been described since 2005 as an etiological agent of respiratory virus infections. From 2001 to 2008 we investigated the etiology of HBoV among adults and children in different groups at risk of presenting complications arising from acute respiratory infection, the investigation was carried out in a tertiary hospital health care system in Brazil. METHODS: HBoV DNA was assayed in 598 respiratory samples from community and hospitalized patients by PCR. RESULTS: Of the 598 tested samples, 2.44% (8/328) of children, including five children with heart disease, and 0.4% (1/270) of adult bone-marrow-transplant were HBoV positive. CONCLUSIONS: These data suggested lower HBoV frequency among different at-risk patients and highlights the need to better understand the real role of HBoV among acute respiratory symptomatic patients. LEAL, E. S. Pós-Dr. Docente da Universidade Federal do Pará, Instituto de Ciências Biológicas, Faculdade de Biotecnologia

INTRODUCTION: Influenza A H1N1 2009 is associated with a high morbidity rate among children around the world, including Brazil. This survey was conducted on samples of symptomatic children (< 12 years) to investigate the influenza virus as the etiological agent of respiratory infections in a day care school in a health facility during the first and second pandemic wave of H1N1 (2009-2010) in São Paulo, Brazil. METHODS: Influenza infections were determined by real-time PCR in 34% (47/137) of children with a median age of 5 years (8 months - 12 years), from June to October 2009 and in 16% (14/85) of those with median age of 6 years (1-12 years), from March to November 2010. RESULTS: In general, most positive cases (64%) occurred in children aged 5-12 years, this age group was significantly the most affected (39.8%, p = 0.001, OR = 8.3, CI 95% 1.9-36.9). Wheezing was reported by 31% (19/61) and dyspnea by 23% (14/61) of the studied patients. An outbreak of influenza H1N1 with an attack rate of 35.7% among children (median age 6 years) was documented in April 2010, before the vaccination campaign against the pandemic virus was extended for children up to 5 years in Brazil. CONCLUSIONS: Therefore, the study reinforces the recommendation to immunize school children to reduce the incidence of the disease. INTRODUÇÃO: Influenza A H1N1 2009 está associado com uma alta taxa de morbidade entre crianças ao redor do mundo, incluindo o Brasil. Esta pesquisa foi realizada em amostras de crianças sintomáticas (< 12 anos) em uma creche escola para filhos de funcionários do hospital durante a primeira e segunda onda pandêmica (2009-2010) em São Paulo, Brasil. MÉTODOS: Infecções pelo vírus influenza foram determinadas por PCR em tempo real em 34% (47/137) em crianças com idade mediana de 5 anos (8 meses -12 anos), entre junho e outubro de 2009 e em 16% (14/85) daquelas com mediana de idade de 6 anos (1-12 anos), de março a novembro de 2010. RESULTADOS: Em geral, a maioria dos casos positivos (64%) ocorreu em crianças com idade entre 5-12 anos, esta faixa etária foi significativamente a mais afetada (39,8%, p = 0,001, OR = 8,3, CI 95%: 1,9-36,9). Chiado foi relatado em 31% (19/61) e dispnéia em 23% (14/61) dos pacientes estudados. Um surto de gripe H1N1 com uma taxa de ataque de 35,7% entre as crianças (mediana de idade de 6 anos) foi documentado em abril de 2010, antes da extensão da campanha de vacinação contra o vírus pandêmico para crianças até 5 anos no Brasil. CONCLUSÕES: Portanto, neste estudo reforça a recomendação para imunizar crianças em idade escolar para reduzir a incidência da doença.

This study assessed the occurrence of human rhinovirus (HRV) species in outpatient children attending day-care in Sao Paulo, Brazil. HRV reverse transcriptase polymerase chain reaction and amplicon sequencing were done in 120 samples collected in 2008. HRV was detected in 27.5% of samples. HRV C was detected in 60.7% of wheezers, a frequency not different from that observed in nonwheezers (69.6%).

A duplex reverse transcriptase polymerase chain reaction-restriction fragment length polymorphism for influenza virus subtyping was applied to 412 patient samples. The assay was able to discriminate all 47 influenza A H1N1 and H3N2 viruses. This rapid technique assessed if positive samples were current circulating strains or an emergent one and could be used as the 1st test in prepandemic stages.

Made available in DSpace on 2013-08-12T18:58:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-01-01 Made available in DSpace on 2013-09-30T18:38:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:51:06Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T13:51:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-01-01 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) Introdução: Infecções agudas do trato respiratório estão entre as doenças mais comuns em todas as pessoas. Os rinovírus têm sido descritos como agente etiológico de mais de 50% das infecções do trato respiratório ao redor do mundo. O objetivo deste trabalho foi avaliar a ocorrência de rinovírus humano (HRV), influenza vírus A e B, vírus respiratório sincicial humano e metapneumovírus (hMPV) em uma população de idosos que apresentava sintomas de gripe ou resfriado, e que residiam na Cidade de Botucatu, Estado de São Paulo, Brasil. Métodos: Foram coletados swabs nasais de 47 idosos após visitas de inclusão, entre os anos de 2002 e 2003 e que foram testadas através de GeneScan RT-PCR. Resultados: HRV foi detectado em 28.6% (14/47) e hMPV em 2% (1/47). De 14 amostras positivas para HRV, 9 foram sequenciadas, mostrando a seguinte distribuição de grupos: grupo A: 6 amostras, grupo B: 1 amostra e grupo C: 2 amostras. Conclusões: A alta incidência de HRV durante os meses de ocorrência de gripe necessita de estudos posteriores para avaliar o impacto desse vírus entre os idosos. A alta frequência de HRV pode contribuir para o aumento da demanda por serviços de saúde durante a estação de influenza. Introduction: Acute respiratory tract infections are the most common illness in all individuals. Rhinoviruses have been reported as the etiology of more than 50% of respiratory tract infections worldwide. The study prospectively evaluated 47 elderly individuals from a group of 384 randomly assigned for acute respiratory viral infections (cold or flu) and assessed the occurrence of human rhinovirus (HRV), influenza A and B, respiratory syncytial virus and metapneumovirus (hMPV) in Botucatu, State of São Paulo, Brazil. Methods: Forty-nine nasal swabs collected from 47 elderly individuals following inclusion visits from 2002 to 2003 were tested by GenScan RT-PCR. HRV-positive samples were sequenced for phylogenetic analysis. Results: No sample was positive for influenza A/B or RSV. HRV was detected in 28.6% (14/47) and hMPV in 2% (1/47). of 14 positive samples, 9 isolates were successfully sequenced, showing the follow group distribution: 6 group A, 1 group B and 2 group C HRVs. Conclusions: The high incidence of HRV during the months of the influenza season requires further study regarding HRV infection impact on respiratory complications among this population. Infection caused by HRV is very frequent and may contribute to increasing the already high demand for healthcare during the influenza season. Univ Fed São Paulo, Dept Med, Infect Dis Unit, Clin Virol Lab, São Paulo, Brazil São Paulo State Univ, Biosci Inst, Human & Anim Virol Lab, Botucatu, SP, Brazil Univ Estadual Campinas, Med Sci Coll, Campinas, SP, Brazil Fed Univ Para, Inst Biotechnol, BR-66059 Belem, Para, Brazil São Paulo State Univ, Biosci Inst, Human & Anim Virol Lab, Botucatu, SP, Brazil