Identification of Human Bocavirus type 4 in a child asymptomatic for respiratory tract infection and acute gastroenteritis - Goiânia, Goiás, Brazil

Abstract Human Bocavirus (HBoV) has been identified from feces and respiratory samples from cases of both acute gastroenteritis and respiratory illness as well as in asymptomatic individuals. The aim of this study was to detect and characterize HBoV from fecal samples collected from hospitalized children aged less than five years old with no symptoms of respiratory tract infection (RTI) or acute gastroenteritis (AGE). The study involved 119 children and one fecal sample was collected from each participant between 2014 and 2015. HBoV was detected using Nested-PCR, and the viral type identified by genomic sequencing. HBoV-4 was identified from one sample obtained from a hospitalized child with soft tissue tumor of the submandibular region. This is the first report of HBoV-4 identification in Brazil, but we consider that this type may be circulating in the country similar to the other types and new investigations are necessary.

Circulating profile of rotavirus a, g and p genotypes before and after vaccine introduction in the Brazilian mid-west 1986-2015

The purpose of this study was to perform a comparative analysis of Rotavirus A (RVA) G and P genotypes circulating in the Brazilian Mid-West in the period 1986-2015. Seven studies conducted from 1986 to 2009 were included, as well as fecal samples obtained in the period 2014-2015. RVA was screened by ELISA and/or PAGE; genotyping by conventional RT-PCR and/or genomic sequencing. A temporal variation in the predominance of G genotypes mainly G1 and G2 with G9 and G12 emergence was observed. Even with vaccination, RVA continues to circulate in the population, requiring continuous virus monitoring

Screening of fecal samples from asymptomatic children, for norovirus detection, using a third generation enzyme immunoassay commercial kit

Norovirus is the leading cause of non-bacterial acute gastroenteritis outbreaks worldwide. Recently, third generation Enzyme Immunoassay (EIA) commercial kits have been developed, and controversial results have been obtained by different studies regarding the sensitivity and specificity of these assays. Therefore, the aim of this study was to test 60 fecal samples, previously tested as positive by RT-PCR for caliciviruses (40 norovirus-positive and 20 sapovirus-positive samples), for qualitative determination of genogroup I and II noroviruses by a commercial EIA kit (RIDASCREEN® Norovirus (C1401) 3rd Generation, R-Biopharm, Darmstadt, Germany). The samples were obtained from 30 children aged less than five years, mostly asymptomatic, who attend a day-care center in Goiânia, Goiás, Brazil. The results conferred a positivity rate for NoV of 35 percentand a specificity rate of 100 percent for the EIA, when compared to the RT-PCR. The test also failed to detect samples that were positive for GI.1 and GI.4 norovirus. The presumably lower viral load of asymptomatic children might be related to the poor sensitivity. Our results reinforce the notion that screening of samples by molecular assays, especially of samples that might have a low number of viral particles such as those obtained from asymptomatic patients, should not be replaced by the use of EIA kits...

Triagem de amostras fecais de crianças assintomáticas utilizando-se um kit comercial de Elisa 3a geração determinação qualitativa de norovírus dos genogrupos I e II por meio de kit comercial de EIE (RIDASCREEN® Norovirus (C1401) 3rd Generation, R-Biopharm, Darmstadt, Germany). Previamente testadas, elas se mostraram positivas para calicivírus por RT-PCR (40 positivas para norovirus e 20 positivas para sapovirus). As amostras foram obtidas de 30 crianças menores de 5 anos de idade, predominantemente assintomáticas, que frequentavam uma creche em Goiânia, Goiás, Brasil. Os resultados revelaram índices de 35 porcento de positividade para os norovírus e de 100 porcento de especificidade para o EIE quando comparado a RT-PCR. O teste também falhou em detectar amostras que eram positivas para norovírus GI.1 e GI.4. A carga viral, presumidamente mais baixa, das crianças assintomáticas pode estar relacionada com a baixa sensibilidade. Os resultados reforçam o entendimento de que a triagem de amostras por ensaios moleculares não deve ser substituída pelo uso de kits de EIE, especialmente quando se tratar de amostras que, presumidamente, apresentem um baixo número de partículas virais como as obtidas de pacientes assintomáticos...

Human bocavirus 1 and 3 infection in children with acute gastroenteritis in Brazil

To determine the positivity rate of human bocavirus (HBoV) 1 and 3 among children who presented with acute gastroenteritis symptoms during the period of 1994-2004 in the Central-West Region of Brazil, 762 faecal samples were tested using polymerase chain reaction (PCR) for the detection of HBoV DNA. Primers for a segment of the non-structural viral protein 1 (NS1) gene of HBoV-1 and HBoV-3 were used. Twelve HBoV-positive samples were further characterised via genomic sequencing and phylogenetic analysis. Of the samples tested, 5.8% (n = 44) were positive for HBoV-1 or HBoV-3 and co-infection was observed in 14 (31.8%) of the 44 HBoV-positive samples. Nine of the 14 samples were also positive for Rotavirus A and five were positive for Aichi virus. The genomic sequencing of the NS1 partial sequence of 12 HBoV-samples showed that 11 samples were characterised as HBoV-1 and that one was characterised as HBoV-3. The phylogenetic analysis showed that the HBoV-1 samples had a high sequence homology to others previously identified in China, Sweden and Brazil. This is the first study conducted in the Central-West Region of Brazil to detect HBoV-1 and HBoV-3 in faecal samples from children with acute gastroenteritis. Further studies are required to define the role of HBoVs as aetiological agents of gastroenteritis.

Monitoring the circulation of rotavirus among children after the introduction of the RotarixTM vaccine in Goiânia, Brazil

The epidemiological features of rotavirus A (RVA) infection differ between children from developing and developed countries which could result in differences in vaccine efficacy around the world. To evaluate the impact of RotarixTM on RVA prevalence, we monitored RVA genotypes circulating in Goiânia by monitoring virus in faecal samples from children that had or had not been previously vaccinated. From February-November of 2008, 220 faecal samples were collected from children in seven day-care centres. RVA detection was performed by two methodologies and the results were confirmed by polyacrylamide gel electrophoresis. From the 220 samples, eight were RVA-positive (3.6 percent) and five were from children that had received either one or two doses of the vaccine. All positive samples were collected from children with diarrhoea during August and September. Genotyping of the RVA characterised five of the viral samples as genotype G2P[4] and one as G8P[4], suggesting that G2P[4] was the predominant circulating genotype in Goiânia during the study. The fact that vaccinated children were also infected by RVA suggests that the vaccine does not fully protect against infection by the G2[P4] RVA genotype.

Gastroenteric virus detection in fecal samples from women in Goiânia, State of Goiás, Brazil / Detecção de vírus gastroentéricos em amostras fecais de mulheres em Goiânia, Estado de Goiás, Brasil

INTRODUCTION: This was a prospective study that included women seen in the obstetrics and gynecology sector of Hospital das Clínicas, Federal University of Goiás, in Goiânia, State of Goiás, with the aim of detecting rotaviruses, adenoviruses, caliciviruses and astroviruses. Eighty-four women participated in the study and from these, 314 fecal samples were collected. Out of all of the women, 29 were seropositive for HIV and 55 were seronegative, and 45 and 39 were pregnant and non-pregnant, respectively. METHODS: Fecal samples were collected from each woman once every two months over the period from July 2006 to June 2007, and they were screened for rotaviruses by means of polyacrylamide gel electrophoresis and immunoenzymatic assays, for caliciviruses and astroviruses by means of RT-PCR and for adenovirus by means of immunoenzymatic assays. The astroviruses were genotyped using nested PCR. RESULTS: Among the 84 patients, 19 (22.6 percent) were positive for either calicivirus (14/19) or astrovirus (6/19), while one women was positive for both viruses in fecal samples collected on different occasions. Most of the positive samples were collected during the months of July and August (astrovirus) and September and October (calicivirus). None of the samples analyzed was positive for rotavirus or adenovirus. Gastroenteric viruses were detected in 13/19 (68.4 percent) of the pregnant women, whether HIV-seropositive or not. CONCLUSIONS: The results from the present study showed that neither pregnancy nor HIV-seropositive status among the women increased the risk of infection by any of the gastroenteric viruses studied. This study presents data on gastroenteric virus detection among pregnant and/or HIV-positive women.

INTRODUÇÃO: Este foi um estudo prospectivo que incluiu mulheres atendidas no setor de obstetrícia e ginecologia do Hospital das Clínicas da Universidade Federal de Goiás, em Goiânia, Estado de Goiás com o objetivo de detectar rotavírus, adenovírus, calicivírus e astrovírus. Oitenta e quatro mulheres participaram no estudo e destas, 314 amostras fecais foram coletadas. Do total de mulheres, 29 eram soropositivas para HIV, 55 soronegativas, 45 e 39 estavam grávidas e não-grávidas, respectivamente. MÉTODOS: Amostras fecais foram coletadas de cada mulher uma vez a cada dois meses pelo período de Julho-2006 a Junho-2007, foram triadas para rotavírus pela metodologia de eletroforese em gel de poliacrilamida (EGPA) e através de ensaio imunoenzimático (EIE), para calicivírus e astrovírus por RT-PCR e por EIE para adenovírus. Os astrovírus foram genotipados por Nested-PCR. RESULTADOS: De 84 pacientes, 19 (22,6 por cento) foram positivas para calicivírus (14/19) ou astrovírus (6/19), sendo que uma mulher foi positiva para ambos os vírus em amostras fecais coletadas em diferentes ocasiões. A maioria das amostras positivas foi coletada no período de Julho a Agosto (astrovírus) e de Setembro a Outubro (calicivírus). Nenhuma das amostras analisadas foi positiva para rotavírus ou adenovírus. Os vírus gastroentéricos foram detectados em 13/19 (68,4 por cento) mulheres grávidas, as quais eram HIV-soropositivas ou não. CONCLUSÕES: Os resultados do presente estudo mostram que nem o estado gravídico das mulheres nem a soropositividade para HIV aumentaram o risco para a infecção por nenhum dos vírus gastroentéricos estudados. Este estudo apresenta dados sobre a detecção de vírus gastroentéricos entre mulheres grávidas e/ou HIV-positivas.

Adenovirus, calicivirus and astrovirus detection in fecal samples of hospitalized children with acute gastroenteritis from Campo Grande, MS, Brazil

We analyzed fecal samples from hospitalized children up to three years of age with acute gastroenteritis at Campo Grande, Mato Grosso do Sul, Brazil, from May 2000-January 2004. Astrovirus and calicivirus were detected by Reverse Transcription-Polymerase Chain Reaction and adenovirus was detected using the Rotavirus and Adenovirus combined immunoenzyme assay. Astrovirus, adenovirus and calicivirus were detected at rates of 3.1 percent, 3.6 percent and 7.6 percent, respectively. These results re-emphasize the need for the establishment of regional vigilance systems to evaluate the impact of enteric viruses on viral gastroenteritis.

Infecções e reinfecções por Rotavirus A: genotipagem e implicações vacinais / Rotavirus A infections and reinfections: genotyping and vaccine implications

OBJETIVOS: Identificar Rotavirus A em crianças com diarréia aguda, determinando os genótipos G e P prevalentes e avaliar a ocorrência de infecções e reinfecções por rotavírus do grupo A em crianças. MÉTODOS: Foram estudadas, prospectivamente, crianças com doença diarréica aguda e identificação de Rotavirus A em Goiânia (GO), durante o período de julho de 2000 a outubro de 2002. Igual número de crianças, pareadas por idade e sexo, que não apresentavam diarréia aguda e sem identificação de rotavírus nas amostras fecais à admissão ao estudo, representou o grupo controle. Foram analisadas a ocorrência de infecções ou reinfecções sintomáticas ou assintomáticas por rotavírus durante o período de estudo, durante um ano de seguimento em ambos os grupos. Todas as amostras positivas foram submetidas a genotipagem G e P através das reações de RT-PCR e Nested PCR. RESULTADOS: A infecção por rotavírus ocorreu em 37,2 por cento (77 de 207 amostras fecais) das crianças com diarréia aguda durante o período do estudo. Os genótipos G e P identificados foram, simultaneamente: G1 (62,3 por cento), G9 (34,4 por cento) e G4 (3,3 por cento) e P[8] (59 por cento), P[6] (7,7 por cento), P[6]+P[8] (23,1 por cento), P[4]+P[8] (7,7 por cento) e P[4]+P[6] (2,6 por cento). As associações de genótipos G e P identificados durante o estudo foram: G1P[8] (77,8 por cento), G9P[8] (11,1 por cento), G4P[8] (5,6 por cento) e G1P[6] (5,6 por cento). Não houve reinfecção por rotavírus nos pacientes do grupo Rotavirus A (+) durante o período de seguimento, enquanto duas crianças do grupo controle apresentaram infecções sintomáticas por rotavírus durante o mesmo período. CONCLUSÕES: Os genótipos G e P predominantes correspondem aos das candidatas atuais à vacina contra rotavírus. Não houve reinfecção por rotavírus pelo período de um ano em relação a todos os genótipos identificados.

Manifestações clínicas e epidemiológicas das infecções por rotavírus A / Clinical and epidemiological trends in rotavirus A infections

Objetivo: analisar as manifestações clínicas e aspectos epidemiológicos relacionados ao rotavirus A. Casusuistica e Métodos: foi realizado estudo prospectivo com crianças de Goiânia, GO, que apresentavam diarréia aguda. Amostras fecais foram avaliadas para rotavírus através de eletroforese em gel e ensaio imunoenzimático...

Epidemiological features of rotavirus infection in Goiânia, Goiás, Brazil, from 1986 to 2000

A total of 2,605 faecal specimens from children up to 10 years old with or without diarrhoea were collected. Samples were obtained from 1986 to 2000 in hospitals, outpatient clinics and day-care centers in Goiânia, Goiás. Two methodologies for viral detection were utilized: a combined enzyme immunoassay for rotavirus and adenovirus and polyacrylamide gel electrophoresis. Results showed 374 (14.4 percent) faecal specimens positive for Rotavirus A, most of them collected from hospitalized children. A significant detection rate of rotavirus during the period from April to August, dry season in Goiânia, and different frequencies of viral detection throughout the years of study were also observed. Rotavirus was significantly related to hospitalization and to diarrhoeal illness in children up to 24 months old. This study reinforces the importance of rotavirus as a cause of diarrhoea in children and may be important in regards to the implementation of rotavirus vaccination strategies in our country