Genotypes of Epstein-Barr virus (EBV1/EBV2) in individuals with infectious mononucleosis in the metropolitan area of Belém, Brazil, between 2005 and 2016

Abstract Two types of Epstein Barr virus (EBV1/EBV2) have been shown to infect humans. Although their genomes are similar, the regions containing the EBNA genes differ. This study aimed to characterize the EBV genotypes of infectious mononucleosis (IM) cases in the metropolitan region of Belém, Brazil, from 2005 to 2016. A total of 8295 suspected cases with symptoms/signs of IM were investigated by infectious disease physicians at Evandro Chagas Institute, Health Care Service, from January 2005 to December 2016. Out of the total, 1645 (19.8%) samples had positive results for EBV by enzyme immunoassay and 251 (15.3%) were submitted to polymerase chain reaction (PCR) technique, using the EBNA3C region, in order to determine the type of EBV. Biochemical testing involving aspartate aminotransferase, alanine aminotransferase and gamma-glutamyl transferase were also performed. EBV type was identified by PCR in 30.3% (76/251) of individuals; of those, 71.1% (54/76) were classified as EBV1, 17.1% (13/76) as EBV2, and 11.8% (9/76) as EBV1+EBV2. The main symptoms/signs observed with EBV1 infection were cervical lymphadenopathy (64.8%, 35/54), fever (63%, 34/54), headache (20.4%, 11/54), arthralgia (20.4%, 11/54), and exanthema (18.5%, 10/54). EBV2 infection was detected in all but two age groups, with an average age of 24 years. The most common signs/symptoms of EBV2 were fever (76.9%, 10/13), average duration of 18 days, and lymphadenopathy (69.2%, 9/13). In contrast, EBV1+EBV2 coinfections were more frequent in those aged five years or less (20.0%, 2/10). The symptoms of EBV1+EBV2 coinfection included fever (66.7%, 6/9), and cervical lymphadenopathy and headache (33.3%, 3/9) each. The mean values of hepatic enzymes according to type of EBV was significantly different (p<0.05) in those EBV1 infected over 14 years of age. Thus, this pioneering study, using molecular methods, identified the EBV genotypes in 30.3% of the samples, with circulation of EBV1, EBV2, and EBV1+EBV2 co-infection in cases of infectious mononucleosis in the northern region of Brazil.

Detection and genotyping of human adenovirus and sapovirus in children with acute gastroenteritis in Belém, Pará, between 1990 and 1992: first detection of GI. 7 and GV. 2 sapoviruses in Brazil

Abstract INTRODUCTION: Acute gastroenteritis (AGE) is one of the most common causes of morbidity and mortality, especially among children from developing countries. Human adenovirus (HAdV) and sapovirus (SaV) are among the agents that cause AGE. The present study aimed to detect and genotype HAdV and SaV in 172 fecal samples from children with AGE, collected during a surveillance study carried out in a low-income community in Belém, Pará, between 1990 and 1992. METHODS: HAdV was detected by nested PCR, using primers Hex1deg/Hex2deg and NeHex3deg/NeHex4deg. SaV was assayed by reverse transcription PCR (RT-PCR), nested PCR, and quantitative PCR. The nucleotide sequence was determined by direct cycle sequencing. RESULTS: Overall, 43% (74/172) of samples were positive for HAdV, of which 70.3% (52/74) were sequenced and classified as belonging to five different species, mostly A and F. For SaV, positivity was 5.2% (9/172) and genotypes GI.1, GI.7, GII.1, and GV.2 were detected. CONCLUSIONS: The present results reinforce the need for further studies to obtain epidemiological data about the circulation of these viruses in Brazil, especially in the Amazon Region, where data from the early 1990's are scarce. Furthermore, the study describes for the first time the detection of SaV genotypes GI.7 and GV.2 in Brazil, showing that these types circulated in the region more than 25 years ago.

High prevalence of norovirus in children with sporadic acute gastroenteritis in Manaus, Amazon Region, northern Brazil

BACKGROUND Norovirus (NoV) is a major cause of acute gastroenteritis (AGE) worldwide, especially in children under five years. Studies involving the detection and molecular characterisation of NoV have been performed in Brazil, demonstrating its importance as an etiological agent of AGE. OBJECTIVES The objectives of this study were to investigate the frequency of human NoV and to genotype the strains isolated from 0-14-year-old patients of AGE in Manaus, Brazil, over a period of two years. METHODS A total of 426 faecal samples were collected between January 2010 and December 2011. All samples were tested for the presence of NoV antigens using a commercial enzyme immunoassay kit. RNA was extracted from all faecal suspensions and reverse transcription-polymerase chain reaction (RT-PCR) for the NoV-polymerase partial region was performed as a trial test. Positive samples were then subjected to PCR with specific primers for partial capsid genes, which were then sequenced. FINDINGS NoV was detected in 150 (35.2%) faecal samples, for at least one of the two techniques used. NoV was detected in children from all age groups, with the highest positivity observed among the group of 1-2 years old. Clinically, fever was verified in 43% of the positive cases and 46.3% of the negative cases, and vomiting was observed in 75.8% and 70.8% cases in these groups, respectively. Monthly distribution showed that the highest positivity was observed in January 2010 (81.2%), followed by February and April 2010 and March 2011, when the positivity rate reached almost 50%. Phylogenetic analyses performed with 65 positive strains demonstrated that 58 (89.2%) cases of NoV belonged to genotype GII.4, five (7.7%) to GII.6, and one (1.5%) each to GII.7 and GII.3. MAIN CONCLUSIONS This research revealed a high circulation of NoV GII.4 in Manaus and contributed to the understanding of the importance of this virus in the aetiology of AGE cases, especially in a region with such few studies available.

Caliciviruses in hospitalized children, São Luís, Maranhão, 1997-1999: detection of norovirus GII. 12

ABSTRACT Gastroenteritis is one of the most common diseases during childhood, with norovirus (NoV) and sapovirus (SaV) being two of its main causes. This study reports for the first time the incidence of these viruses in hospitalized children with and without gastroenteritis in São Luís, Maranhão. A total of 136 fecal samples were tested by enzyme immunoassays (EIA) for the detection of NoV and by reverse transcription-polymerase chain reaction (RT-PCR) for detection of both NoV and SaV. Positive samples for both agents were subjected to sequencing. The overall frequency of NoV as detected by EIA and RT-PCR was 17.6% (24/136) and 32.6% (15/46), respectively in diarrheic patients and 10.0% (9/90) in non-diarrheic patients (p < 0.01). Of the diarrheic patients, 17% had fever, vomiting and anorexia, and 13% developed fever, vomiting and abdominal pain. Of the 24 NoV-positive samples, 50% (12/24) were sequenced and classified as genotypes GII.3 (n = 1), GII.4 (6), GII.5 (1), GII.7 (2), GII.12 (1) and GII.16 (1). SaV frequency was 9.8% (11/112), with 22.6% (7/31) in diarrheic patients and 4.9% (4/81) in nondiarrheic (p = 0.04) ones. In diarrheic cases, 27.3% had fever, vomiting and anorexia, whereas 18.2% had fever, anorexia and abdominal pain. One SaV-positive sample was sequenced and classified as GII.1. These results show a high genetic diversity of NoV and higher prevalence of NoV compared to SaV. Our data highlight the importance of NoV and SaV as enteropathogens in São Luís, Maranhão.

The prevalence of norovirus, astrovirus and adenovirus infections among hospitalised children with acute gastroenteritis in Porto Velho, state of Rondônia, western Brazilian Amazon

Although viruses are well-established causes of acute gastroenteritis, few data on the circulation of these pathogens in Porto Velho, state of Rondônia, Brazil, are available. Thus, faecal samples from hospitalised diarrhoeic children, under six years of age, were collected and tested for the presence of norovirus (NoV), adenovirus (AdV) and astrovirus (AstV) from February 2010-February 2012. Specimens were screened by reverse-transcription polymerase chain reaction and viruses were found in 10.7% (63/591) of the cases. NoV, AdV and AstV were detected in 7.8%, 2% and 0.8% of the samples, respectively. NoV infection was observed at all ages and was most prevalent in zero-18-month-old children (84.7%; p = 0.002). A higher incidence of NoV was detected from February-April 2010, when it was found in 52.2% of the cases. Co-infections involving these viruses, rotavirus and enteropathogenic bacteria were detected in 44.4% (28/63) of the children with viral diarrhoea. Nosocomial infections were demonstrated in 28.6% (18/63) of the cases in which viruses were detected. The present paper reports, for the first time, the circulation of NoV and AstV among the paediatric population of Porto Velho and it contributes to our understanding of the roles of these pathogens in gastrointestinal infections.

Enteropathogensddetected in a daycare center, sutheastern Brazil: bacteria, viirus, and parasite research / Enteropatógenos detectados em crianças de creche no Sudeste do Brasil: pesquisa de bactérias, vírus e parasitos

Introduction: The objective of this study was to determine the prevalence and etiological profile of enteropathogens in children from a daycare center. Methods: From October 2010 to February 2011 stool samples from 100 children enrolled in a government daycare center in the municipality of São José do Rio Preto, in the state of São Paulo, were collected and analyzed. Results: A total of 246 bacteria were isolated in 99% of the fecal samples; 129 were in the diarrheal group and 117 in the non-diarrheal group. Seventy-three strains of Escherichia coli were isolated, 19 of Enterobacter, one of Alcaligenes and one of Proteus. There were 14 cases of mixed colonization with Enterobacter and E. coli. Norovirus and Astrovirus were detected in children with clinical signs suggestive of diarrhea. These viruses were detected exclusively among children residing in urban areas. All fecal samples were negative for the presence of the rotavirus species A and C. The presence of Giardia lamblia, Entamoeba coli, Endolimax nana and hookworm was observed. A significant association was found between food consumption outside home and daycare center and the presence of intestinal parasites. Conclusions: For children of this daycare center, intestinal infection due to pathogens does not seem to have contributed to the occurrence of diarrhea or other intestinal symptoms. The observed differences may be due to the wide diversity of geographical, social and economic characteristics and the climate of Brazil, all of which have been reported as critical factors in the modulation of the frequency of different enteropathogens.

Introdução: O objetivo deste estudo foi determinar a prevalência e o perfil etiológico de enteropatógenos em crianças de uma creche. Métodos: No período de outubro de 2010 a fevereiro de 2011 foram coletadas e analisadas amostras de fezes de 100 crianças matriculadas em creche do governo no município de São José do Rio Preto, Estado de São Paulo. Resultados: Um total de 246 bactérias foram isoladas em 99% das amostras de fezes; 129 eram diarreicas e 117 não-diarreicas. Foram isoladas setenta e três cepas de Escherichia coli, 19 de Enterobacter, uma de Alcaligenes e uma de Proteus. Foram detectados 14 casos de colonização mista com Enterobacter e de E. coli. Norovírus e Astrovirus foram detectados em crianças com sinais clínicos sugestivos de diarréia. Estes vírus foram detectados exclusivamente entre as crianças residentes em áreas urbanas. Todas as amostras fecais foram negativas para a presença das espécies de rotavírus A e C. Foi observada a presença de Giardia lamblia, Entamoeba coli, Endolimax nana e ancilostomídeos. Foi encontrada associação significativa entre o consumo de alimentos fora do centro da casa e creche e a presença de parasitos intestinais. Conclusões: Para as crianças desta creche, a infecção intestinal por patógenos não parece ter contribuido para a ocorrência de diarreia ou outros sintomas intestinais. As diferenças observadas podem ser atribuídas à grande diversidade de características geográficas, sociais e econômicas e o clima do Brasil, as quais tem sido relatadas como fatores críticos para a modulação da frequência de diferentes enteropatógenos.

Group A rotavirus and norovirus display sharply distinct seasonal profiles in Belém, northern Brazil

Several viruses have been associated with acute gastroenteritis (AGE), and group A rotavirus (RVA) and nor-ovirus (NoV) are the most prevalent. This study aimed to assess their prevalence among children hospitalised for diarrhoea during a three-year surveillance study. From May 2008-April 2011, overall positivity rates of 21.6% (628/2904) and 35.4% (171/483) were observed for RVA and NoV, respectively. The seasonality observed indicated distinct patterns when both viruses were compared. This finding may explain why hospitalisation for AGE remains constant throughout the year. Continuous AGE monitoring is needed to better assess the patterns of infection.

Detecção de rotavírus por imunocromatografia: ensaio simples e de rápida execução / Rotavírus detection by imunocromatografic: simple assay with a fast execution

Introdução:As gastroenterites agudas são responsáveis por uma alta taxa de morbidade e mortalidade entre crianças em todo mundo, sendo que os rotavírus constituem os principais agentes etiológicos de diarréia, causando a morte de 418.000 a 520.000 crianças ao ano, principalmente nos países em desenvolvimento. Objetivo:Descrever os dados obtidos com um novo "kit" de diagnóstico para detecção de rotavírus demonstrando sua especificidade e sensibilidade. Método: Noventa suspensões fecais foram testadas, simultaneamente, quanto à presença de rotavírus usando o "kit" Rota Strip, do Laboratório Coris BioConcept, e comparativamente o ELISA comercial Premier Rotaclone, Meridian Bioscience Inc. Ohio, USA. Resultados:Ambas as técnicas demonstraram sensibilidade e especificidade comparáveis, considerando que nenhum falso positivo nem negativo foi observado. Conclusão:Nossos resultados sugerem que o Rota strip é um novo e prático instrumento de diagnóstico, de rápida execução e que poderá ser util quando usado na prática clínica no hos´pital e laboratórios de saúde pública

Diagnóstico de amebíase intestinal utilizando métodos coproscópicos e imunológicos em amostra da populaçäo da área metropolitana de Belém, Pará, Brasil / Diagnosis of intestinal amebiasis using coproscopic and immunological methods in a population sample in Greater Metropolitan Belém, Pará, Brazil

Expöe a comparaçäo de métodos de diagnóstico de Entamoeba histolytica em amostra da populaçäo de Belém do Pará. Foram analisadas amostras fecais de crianças e adultos (Grupo I), amostras fecais e soros de adultos (Grupo II) e material fecal de crianças (Grupo III). Nos grupos I e III foram empregados os métodos direto com lugol (MD), Faust e cols. (MFF) e ELISA (detecçäo de coproantígeno anti-GIAP de E. histolytica); no grupo II, MD, hematoxilina férrica (HF), MFF, ELISA e reaçäo de imunofluorescência indireta (RIFI) para detecçäo de anticorpos IgG. A positividade encontrada foi de 10,50 por cento, empregando (MD + MFF) e de 28,99 por cento pelo ELISA. Näo houve correlaçäo entre positividade e grupo etário. No Grupo II (n = 87), a positividade encontrada foi de 4,59 por cento pelos métodos coproscópicos (MD + MFF), 8,04 por cento por HF, 4,59 por cento pela RIFI e 21,83 por cento pelo ELISA. O teste de ELISA foi o mais sensível para todos os grupos. Conclui-se que a RIFI ainda näo é ferramenta útil para diagnóstico desta protozoose. O teste de ELISA, de fácil execuçäo, foi feito em 1/3 do tempo usado para HF e RIFI, melhorando a qualidade do diagnóstico. Recomenda-se o ELISA como método de diagnóstico nos casos suspeitos de infecçöes com E.histolytica.

Três infecçöes sucessivas por rotavírus em criança de Belém, Pará, Brasil / Three successive rotavírus infections ina child living in Belém, Pará, Brasil

Três infecçöes, subsequentes, em uma única criança foram observadas ao longo de um estudo prospectivo na àrea urbana de Belém, Pará, Brasil, no decorrer de seus primeiros vinte e sete meses de vida. A detecçäo de rotavírus classificado como subgrupo II, sorotipo näo determinado, foi obtido nas duas primeiras infecçöes, sucedendo-se uma terceira pelo subgrupo I, sorotipo 2. Nesses episódios, observaram-se dois perfis eletroforéticos diferentes. A primeira infecçäo ocorreu aos 4 meses e as seguintes no 20§ e 27§, respectivamente. os processos infecciosos ocorreram de forma sintomática, sendo que em dois dos três episódios, as manifestaçöes clínicas forma mais severas, o segundo, em maior grau que o primeiro e, o terceiro, mais benigno que os anteriores