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.

Detection of caliciviruses associated with acute infantile gastroenteritis in Salvador, an urban center in Northeast Brazil

Acute gastroenteritis caused by viruses is one of the leading causes of infantile morbidity. The aim of the present study was to investigate the presence of human caliciviruses of the genera norovirus and sapovirus in children up to 3 years of age with acute gastroenteritis from low-income communities in the city of Salvador, Brazil. This study is an extension of previous work carried out to establish the profile of the most prevalent enteric pathogens present in these communities. In this report, 139 fecal samples, collected from July 2001 to January 2002 were analyzed by RT-PCR and 13 (9 percent) were positive for human caliciviruses. By sequencing, seven isolates were characterized as norovirus genogroup GII and one as sapovirus genotype GII/1. Sequencing of the previously detected group-A rotaviruses and human astroviruses was also performed and revealed the circulation of rotavirus group A genotypes G1P[8] and G9P[8], and human astrovirus genotypes 6, 7, and 8. No mixed infection was observed. Community-based studies provide geographically representative information on disease burden. However, there are only a few reports in developing countries concerning the genotypes of the most important gastroenteric viruses detected in such communities. The present findings demonstrate the wide diversity of genotypes of the most important viruses responsible for acute gastroenteritis circulating in low-income communities.

Caracterización molecular de calicivirus aislados de brotes de gastroenteritis ocurridos en la Argentina durante los años 2005 y 2006 / Molecular characterization of calicivirus strains detected in outbreaks of gastroenteritis occurring in Argentina during 2005 and 2006

Con el objetivo de determinar la incidencia de calicivirus, rotavirus y astrovirus en brotes de gastroenteritis ocurridos en diversas regiones de la Argentina durante los años 2005 y 2006, se analizaron muestras de materia fecal provenientes de 7 brotes con resultado de coprocultivo negativo. Para el diagnóstico de rotavirus se utilizó un ELISA comercial, mientras que para el diagnóstico de calicivirus y astrovirus se utilizó el método de RT-PCR. De las 74 muestras analizadas, 20 fueron positivas para calicivirus, 17 para rotavirus y una para astrovirus. No se identificaron infecciones virales mixtas. En 5 muestras positivas para calicivirus se secuenció una región del gen de la polimerasa; 4 de ellas correspondieron al género Norovirus y una al género Sapovirus. El análisis filogenético de las muestras secuenciadas determinó la presencia de norovirus de los genogrupos GI y GII; dentro de este último, se identificaron los genotipos GII-4, GII-b y GII-17. El análisis de la muestra en la cual se identificó sapovirus reveló la presencia del genotipo GI-1. Este estudio representa una continuación del análisis epidemiológico molecular de calicivirus asociados a brotes de gastroenteritis iniciado en 2004 y constituye la primera comunicación de la circulación de norovirus del genotipo GII-17 en la Argentina.

In order to determine the incidence of calicivirus, rotavirus and astrovirus in outbreaks of gastroenteritis occurring in different regions of Argentina during 2005 and 2006, fecal samples from seven nonbacterial outbreaks were analyzed. A commercial ELISA was used for rotavirus detection, while RT-PCRs were used for calicivirus and astrovirus. Of the 74 samples analyzed, 20 were calicivirus positive, 17 were rotavirus positive and one was astrovirus positive. No mixed infections were detected. A partial region of the RdRp gene was sequenced in five calicivirus positive-samples; 4 of them belonged to Norovirus genus and one to Sapovirus genus. The phylogenetic analysis of norovirus-positive-samples revealed the presence of strains from genogroups GI and GII; genotypes GII- 4, GII-b and GII-17 were identified within the latter. Phylogenetic the sapovirus-positive-sample revealed the presence of genotype GI-1. This study represents a follow-up of the of molecular epidemiology analysis of calicivirus associated to gastroenteritis outbreaks that have been carried out by our group since 2004, and constitutes the first report of the circulation of genotype GII-17 in Argentina.

Detection and phylogenetic analysis of porcine enteric calicivirus, genetically related to the Cowden strain of sapovirus genogroup III, in Brazilian swine herds

Sapovirus of the Caliciviridae family is an important agent of acute gastroenteritis in children and piglets. The Sapovirus genus is divided into seven genogroups (G), and strains from the GIII, GVI and GVII are associated with infections in swine. Despite the high prevalence in some countries, there are no studies related to the presence of porcine enteric sapovirus infections in piglets in Brazil. In the present study, 18 fecal specimens from piglets up to 28 days were examined to determine the presence of sapovirus genome by RT-PCR assay, using primers designed to amplify a 331 bp segment of the RNA polymerase gene. In 44.4 percent (8/18) of fecal samples, an amplified DNA fragment was obtained. One of these fragments was sequenced and submitted to molecular and phylogenetic analysis. This analysis revealed high similarity, with nucleotides (87 percent) and amino acids (97.8 percent), to the Cowden strain, the GIII prototype of porcine enteric calicivirus. This is the first description of sapovirus in Brazilian swine herds.

O sapovírus classificado na família Caliciviridae é um importante causador de gastroenterite aguda em crianças e leitões. O gênero Sapovirus é dividido em sete genogrupos (G), sendo que as estirpes dos GIII, GVI e GVII estão associadas com infecção em suínos. Apesar da alta prevalência da infecção em alguns países, ainda não existem estudos referentes à presença do calicivírus entérico suíno nos rebanhos brasileiros. No presente estudo 18 amostras de fezes de leitões com até 28 dias foram avaliadas pela RT-PCR para a presença do genoma do sapovírus, utilizando os primers desenvolvidos para amplificar um segmento de 331 pb do gene da RNA polimerase viral. Em 44,4 por cento (8/18) das amostras foi amplificado um fragmento de DNA. Um desses amplicons foi seqüenciado e pela análise molecular e filogenética foi verificada similaridade de 87 por cento em nucleotídeos e 97,8 por cento em aminoácidos com a estirpe Cowden, protótipo do GIII. Esta é a primeira descrição do sapovírus em rebanhos suínos brasileiros.