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.

Molecular characterization of hepatitis A virus isolates from Goiânia, Gioás, Brazil

Hepatitis A virus (HAV) infection is a public health problem worldwide and the virus has been classified into six genotypes. In Brazil, the only genotype that has been found is genotype I, predominately from subgenotype IA. Here, the HAV genotypes were analyzed of 18 isolates circulating between 1996-2001 in Goiânia, state of Goiás, Brazil. Viral RNA was extracted from 18 serum samples and amplified (RT-PCR/nested-PCR), followed by the genomic sequencing of the VP1/2A junction region of the HAV genome. Sequences of 168 nucleotides were compared and analyzed using the BLAST N, Clustal X and PAUP v. 4.10b programs. All samples were classified as genotype I, with 10 belonging to subgenotype IA and eight to subgenotype IB. The subgenotype IA isolates showed greater diversity than the subgenotype IB isolates at the nucleotide level. Elevated identity values were found between isolates obtained in this study and those from other regions of the world, including Brazil, highlighting the high conservation among different isolates of this virus. However, changes in the HAV subgenotype circulation could also be observed during the evaluated period.

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.

Rotavírus A em crianças de até três anos de idade, hospitalizadas com gastroenterite aguda em Campo Grande, Estado do Mato Grosso do Sul / Rotavirus A among hospitalized infants, up to three years of age, with acute gastroenteritis in Campo Grande, State of Mato Grosso do Sul

Através da eletroforese em gel de poliacrilamida e do ensaio imunenzimático combinado para rotavírus e adenovirus, foram analisadas 380 amostras fecais de crianças com até 3 anos, hospitalizadas com diarréia aguda, entre maio de 2000 e janeiro de 2004, em Campo Grande, MS. Do total de amostras, 88 (23,2 por cento) foram positivas para Rotavirus A. Dentre essas, 81 (92 por cento) tiveram padrão eletroferotípico definido, sendo 77 (87,5 por cento) de padrão longo e quatro (4,5 por cento) de padrão curto. A caracterização genotípica G e P foi feita por RT-Nested-PCR para 85 amostras, sendo 56 (65,9 por cento) genotipáveis para genótipo G. Dentre essas, 49 (87,5 por cento) foram G1, cinco (8,9 por cento) G4, uma (1,8 por cento) G3 e uma (1,8 por cento) G9. Considerando a genotipagem P, 37 (43,5 por cento) foram genotipáveis e todas eram P[8]. A associação G e P mais observada foi G1P[8], 33 (89,2 por cento) amostras; seguida de G4P[8], duas (5,4 por cento) amostras; G3P[8], uma (2,7 por cento) amostra; e G9P[8], uma (2,7 por cento) amostra.

Polyacrylamide gel electrophoresis and combined immunoenzyme assay for rotavirus and adenovirus were used to analyze 380 fecal samples from children up to three years of age who were hospitalized with acute diarrhea in Campo Grande, State of Mato Grosso do Sul, between May 2000 and January 2004. Among all the samples, 88 (23. 2 percent) were positive for Rotavirus A. Out of these, 81 (92 percent) had a defined electrophoretic pattern: 77 (87. 5 percent) with a long pattern and four (4. 5 percent) with a short pattern. Genotype G and P characterization was done by nested RT-PCR for 85 samples, of which 56 (65. 9 percent) were genotyped as type G. Among these, 49 (87. 5 percent) were G1, five (8. 9 percent) were G4, one (1. 8 percent) was G3 and one (1. 8 percent) was G9. The genotype was found to be type P in 37 samples (43. 5 percent) and all of these were P[8]. The G and P association most observed was G1P[8], with 33 samples (89. 2 percent), followed by G4P[8], two samples (5. 4 percent); G3P[8], one sample (2. 7 percent); and G9P[8], one sample (2. 7 percent).

Astrovirus infection in children living in the Central West region of Brazil

This study presents data regarding the circulation of astrovirus in Goiânia-GO and Brasília-DF. These viruses were detected in fecal samples from hospitalized children up to five years old with and without acute gastroenteritis. A total of 1244 fecal samples were collected in two periods, 1994 to 1996 (Brasília) and 1998 to 2002 (Goiânia and Brasília), and were analyzed for viral RNA using the reverse transcription-polymerase chain reaction (RT-PCR). Positivity rates of 4.3 and 0.5 percent for astrovirus were observed in children with acute gastroenteritis and those without gastroenteritis, respectively. Among children with gastroenteritis no statistically significant difference was seen with regards to viral positivity rates in relation to gender and age. However, a higher incidence rate was observed for children from Brasília aged 36 months or more. Overall, astroviruses occurred predominantly from September to March in the two cities, suggesting a seasonal pattern for these viruses which coincides with the highest relative air humidity period. The results of this study highlight the importance of astrovirus as an etiologic agent of acute gastroenteritis in children of the Central West region of Brazil.

Detection of calicivirus from fecal samples from children with acute gastroenteritis in the West Central region of Brazil

The objective of this study was to describe the circulation of caliciviruses in the West Central region of Brazil and its correlation with children's gender and age, as well as with the year and months of the sample collection. Reverse transcriptase-polymerase chain reaction was performed to detect the human calicivirus genome in 1006 fecal samples that were collected in Goiânia (n = 696) and Brasília (n = 310). Viral RNA was detected in 8.6 percent of the samples. No significant difference in viral prevalence was found regarding gender, age or year of the sample. However, it was observed that in Goiânia, there is a higher incidence of caliciviruses from September to March. The analysis employing three primer pairs demonstrated that the Ni/E3 or JV12/13 primer pairs, which detect norovirus (NoV), detected 41 positive samples while the 289/290 primer pair, which detects NoV or sapovirus, detected the remaining 46 samples. Calicivirus circulates in the West Central region of Brazil and for better detection of this virus it is important to use more than one primer pair. Also, we conclude that the seasonality presented by this virus is related to higher humidity in the period.

Prevalence of hepatitis A virus infection in Goiânia, Goiás, Brazil, by molecular and serological procedures, 1995-2002

In this study, a total of 865 serum samples were collected between 1995 and 2002 from individuals living in Goiânia, Central Brazil, and clinically suspected of hepatitis. After exclusion of 162 samples which were positive for hepatitis B virus or hepatitis C virus, 703 samples were tested for anti-hepatitis A virus (anti-HAV) IgM antibodies by enzyme immunoassay. In addition, 588 of these samples and 22 fecal samples were analyzed by reverse transcription-nested PCR for HAV RNA detection, with positivity indices of 13.1 percent (77/588) and 54.5 percent (12/22), respectively. A similar index of viral RNA detection in anti-HAV-IgM positive or negative samples was observed in serum samples. HAV infection is a public health problem worldwide and this study underscores the extent of HAV circulation in our region.

Seroprevalence of hepatitis B virus infection and seroconvertion to anti-HBsAg in laboratory staff in Goiânia, Goiás

Neste estudo, foram analisadas 648 amostras de soro, provenientes de profissionais de laboratório de Goiânia-Goiás, visando a detecção de três marcadores sorológicos do VHB: HBsAg, anti-HBsAg e anti-HBcAg. As amostras HBsAg e anti-HBcAg positivas foram também analisadas para os marcadores HBeAg, anti-HBeAg e anti-HBcAgIgM. Foi observado um percentual de soropositividade para infecção pelo VHB de 24,1% sendo que 0,7% destes foram positivos para o HBsAg. O DNA viral foi detectado por PCR nas duas amostras HBsAg positivas. Dos 648 profissionais, 74,5% relataram vacinação para o VHB e, destes, 89,9% apresentaram soroconversão. Trabalho direto com fluidos biológicos bem como trabalho em serviços gerais representaram risco significativo para aquisição da infecção pelo VHB. Os resultados do estudo mostram que embora tenha havido um aumento no índice de vacinação entre os trabalhadores de laboratório, o índice de infecção pelo VHB manteve-se inalterado em relação ao tempo na região.

Calicivírus humanos / Human caliciviruses

Os calicivírus humanos (HuCVs) têm sido descritos como importantes patógenos nas gastrenterites agudas em todo o mundo. Esses vírus pertencem à família Caliciviridae, que possui quatro gêneros, dentre os quais apenas Norovirus e Sapovirus infectam o homem. Possuem RNA de fita simples, de polaridade positiva e são poliadenilados na extremidade 3'.São desnudos, apresentam um nucleocapsídeo de simetria icosaédrica, têm diâmetro de 27-40 nm e uma arquitetura típica com cerca de 32 depressões em formato de cálice. Os HuCVs possuem uma grande diversidade genética, o que dificulta estudos de caracterização genômica. O conhecimento sobre a epidemiologia da infecção por calicivírus e a disponibilidade de métodos rápidos e eficazes de detecção desses vírus são importantes para as estratégias de prevenção e controle.

Soroprevalência da infecção pelo vírus da hepatite B em portadores de doença mental / Seroprevalence of hepatitis B virus infection in patients with mental problems

OBJETIVOS: O vírus da hepatite B representa um importante problema de saúde pública mundial e é responsável por altos índices de morbidade e mortalidade. Este estudo objetivou a determinação da soroprevalência da infecção pelo vírus da hepatite B (VHB) em pacientes internados em instituições psiquiátricas e em indivíduos com síndrome de Down, a detecção do DNA viral nas amostras positivas para HBsAg e anti-HBc total, e a caracterização dos subtipos do HBsAg. MÉTODOS: O estudo avaliou 433 indivíduos, sendo que 408 eram portadores de doença mental e internos em instituições psiquiátricas (71 deles tinham também dependência química) e 25 eram indivíduos com síndrome de Down, não institucionalizados. Amostras sangüíneas de todos os participantes foram testadas para os marcadores do VHB: HBsAg, anti-HBs e anti-HBc total pelo ensaio imunoenzimático (ELISA). Soros positivos ao HBsAg foram testados também para o anti-HBc IgM, HBeAg, anti-HBe, e subtipadas pela técnica de imunodifusão radial. A detecção do DNA-VHB foi realizada em amostras reagentes ao HBsAg e/ou anti-HBc total, utilizando-se a metodologia da reação em cadeia pela polimerase (PCR). RESULTADOS: Foi observado um índice global de soropositividade para o VHB de 22,4 por cento, sendo que 1,6 por cento dos indivíduos foram positivos para HBsAg. Dentre as amostras HBsAg positivas, cinco foram subtipáveis pela metodologia de imunodifusão radial, tendo sido observados os subtipos adw2, adw4 e ayw3. O DNA viral foi detectado, pela metodologia da PCR, em três amostras positivas para o HBsAg e em 11 amostras positivas para anti-HBc total/anti-HBs. No grupo de indivíduos com problema mental, com ou sem dependência química (N=408), a admissão hospitalar múltipla (reinternação) foi considerada fator de risco para infecção pelo VHB. CONCLUSÃO: Estes resultados indicam alta prevalência do VHB nesta população e reforçam a necessidade de medidas profiláticas específicas para reduzir o risco de infecção entre indivíduos com distúrbio mental e retardo mental.