Estudo epidemiológico da diarreia aguda, correlação com etiologia viral e antígenos do grupo histo-sanguíneo em crianças ≤ 5 anos atendidas no Hospital da Criança de Santo Antônio em Boa Vista, Roraima, 2016-2017 / Epidemiological study of acute diarrhea, correlation with viral etiology and antigens of the histo-blood group in children ≤ 5 years old attended at the Hospital da Criança of Santo Antônio in Boa Vista, Roraima, 2016-2017

Neste estudo avaliamos o impacto das doenças diarreicas agudas (DDA) associadas aos rotavírus A (RVA), norovírus, adenovírus (HAdV), sapovírus (SaV), bocavírus (HBoV) e astrovírus (HAstV) em crianças ≤ 5 anos atendidas na emergência do Hospital da Criança de Santo Antônio (HCSA), Boa Vista, Roraima (RR), no período de outubro de 2016 a outubro de 2017. Foram coletadas, paralelamente, amostras de fezes e de saliva de 734 crianças sendo 485 com DDA e 249 com infecção respiratória aguda (IRA, grupo controle). O estudo teve a aprovação do CEP:1.333.480, 23/11/2015, UFRR. Os RVA, norovírus, HAdV, SaV e HBoV foram pesquisados nas 734 amostras de fezes e HAstV em 170 amostras de crianças com DDA. Adicionalmente, a presença dos HBoV foi também investigada em 38 amostras de saliva sendo 25 de crianças com DDA e 13 IRA. O perfil de susceptibilidade AB0, Lewis e secretor dos antígenos do grupo histosanguíneo (HBGA) foi determinado para todas as 734 crianças em salivas pela fenotipagem e em amostras de salivas pela genotipagem, sendo para o gene FUT2 em 166 amostras e para FUT3 em 42 amostras. Os aspectos clínicos, epidemiológicos e a cobertura da vacina Rotarix® (RV1) foram também avaliados. Os RVA, norovírus e SaV foram investigados pela metodologia de transcrição reversa seguida de amplificação genômica quantitativa (RT-qPCR); os HBoV e HAdV pela amplificação genômica quantitativa (qPCR) e os HAstV por amplificação genômica qualitativa (RT-PCR). A genotipagem dos HBoV e a detecção/genotipagem dos HAstV foi realizada pela PCR seguida de sequenciamento nucleotídico (método Sanger). O Ensaio imunoenzimático (EIA) e a amplificação genômica específica (PCR-touchdown), seguida de sequenciamento nucleotídico (Sanger) foram utilizadas respectivamente para a fenotipagem e genotipagem dos HBGA.

Nas crianças com DDA (n=485), observou-se as seguintes frequências virais: RVA (22,7%), norovírus (38%), HAdV (33,6%), SaV (7,3%) e HBoV (14,2%). Nas crianças com IRA (n=249), observou as seguintes frequências: RVA (19,3%), norovírus (21,3%), HAdV (39,5%), SaV (5,6%) e HBoV (14,1%). O perfil de detecção do HBoV nas 76 amostras de saliva e fezes pareadas foi diferente e correlacionado com os genótipos 1 a 3 detectados. Quanto aos HAstV, nas 170 amostras investigadas, observou-se as seguintes frequências e genótipos: HAstV clássicos: HAstV3 (0,60%) e HAsV5 (1,8%); HAstV não clássicos: HAstVMLB1-2 (3,5%), sendo esta a primeira descrição do HAstVMLB2 no Brasil. A cobertura vacinal para RV1 calculada foi de 61% e crianças vacinadas na faixa etária entre 6 e 24 meses apresentaram frequências mais elevadas de infecção pelo RVA. As 734 crianças apresentaram majoritariamente (54.5%) o perfil Lea+b+ (fraco secretor) e polimorfirmos de nucleotídico único (SNPs) foram detectados nos genes FUT2 e FUT3. Foi detectada susceptibilidade (HBGA) para a infecção pelos HAdV em crianças com IRA, perfil fraco secretor e grupo sanguíneo A ou O, de acordo com valores de Odds Ratio (OR) calculado. As frequência dos vírus detectados principalmente para os norovírus e HAdV em crianças com DDA, demostram a importância da etiologia viral nas DDA em crianças ≤ 5 anos de idade no período do estudo e podem estar relacionadas ao fator de susceptibilidade ao HBGA (incluindo heterogeneidade genética) e a baixa cobertura de RV1. (AU)

Padronização, implantação e caracterização molecular de sapovírus em amostras de pacientes com gastroenterites / Standardization, implantation and molecular characterization of sapoviruses in samples from patients with gastroenteritis

No mundo são registrados 1,7 bilhões de casos de doença diarreica aguda (DDA) por ano, sendo considerada a segunda maior causa de morte em crianças menores de 5 anos de idade. Após a introdução da vacina contra rotavírus (RVA) em diversos países do mundo e o aprimoramento das técnicas de diagnóstico molecular, outros vírus emergiram como importantes causadores de DDA, como os sapovírus (SaV), sendo frequentemente associados à surtos e casos esporádicos em adultos e crianças. No Brasil, pouco se sabe sobre a ocorrência do SaV e seu impacto em Saúde Pública. O presente estudo teve por objetivo a padronização, implantação e caracterização molecular de SaV em amostras de fezes de pacientes com gastroenterite. Foram selecionadas amostras negativas para RVA, norovírus e adenovírus entéricos, totalizando 3974 amostras coletadas de pacientes de 0 a 91 anos, provenientes das regiões Sudeste, Sul e Centro-Oeste do Brasil. Os ensaios de PCR convencional e rRT-PCR foram padronizados para implantação no diagnóstico laboratorial dos SaV. Os SaV foram detectados em 149 (3,7%) das amostras analisadas: a faixa etária de 0 a 2 anos foi a mais acometida (5,7%). Este foi o primeiro estudo a detectar SaV no estado de SP, com detecção expressiva no ano de 2016 (8,9%). A variabilidade genotípica dos SaV nas amostras estudadas foi detectada após a análise de sequencia parcial da VP1: genogrupo GI (GI.1, GI.2, GI.3, GI.6), genogrupo GII (GII.1, GII.2, GII.3, GII.4 GII.5) e genogrupo GIV (GIV.1).

Human sapovirus GI. 2 and GI. 3 from children with acute gastroenteritis in northern Brazil

Human sapoviruses (HSaV) are considered important causative agents of acute gastroenteritis in humans worldwide. However, knowledge of the genetic characteristics of the whole genome of HSaV in Brazil is limited. Here we report the complete genome sequences of six HSaVs GI.2 and two GI.3 strains obtained from children with acute gastroenteritis in the Northern region of Brazil. Next generation sequencing was used to obtain the full genome and molecular characterization of the genome was performed. Phylogenetic analysis of the genome was also performed. Only one complete HSaV GI.2 genome characterization in the country precedes that of the present study. This is the first complete genome sequence of genotype GI.3 in Brazil. The data obtained in this investigation can contribute to the augmentation of the database on the molecular diversity of HSaVs strains circulating in Brazil, and to the improvement of current typing protocols.

Evaluation of a New Multiplex Real-Time PCR Assay for Detecting Gastroenteritis-Causing Viruses in Stool Samples

BACKGROUND: Diarrhea has been the second leading cause of death among children under the age of five, and the rapid and accurate pathogen diagnosis in patients with diarrhea is crucial for reducing morbidity and mortality. A newly developed one-step multiplex real-time PCR assay, the Allplex GI-Virus Assay, was evaluated for its ability to detect six diarrhea-causing viruses (rotavirus, norovirus genogroup I (GI) and genogroup II (GII), enteric adenovirus, astrovirus, and sapovirus) in stool samples. METHODS: The performance of the Allplex assay was compared with those of another multiplex PCR assay (Seeplex Diarrhea-V Ace Detection) and genotyping by sequencing, using 446 stool samples from patients with acute gastroenteritis. RESULTS: The overall agreement rates between the results of the Allplex and Seeplex assays were 98.7% for rotavirus, 99.1% for norovirus GI, 93.3% for norovirus GII, 98.0% for adenovirus, and 99.6% for astrovirus. The overall agreement rates between the Allplex assay and genotyping were 99.1% for rotavirus, 99.1% for norovirus GI, 98.7% for norovirus GII, 89.7% for adenovirus, 98.2% for astrovirus, and 99.8% for sapovirus. In addition, eight rotavirus genotypes, three norovirus GI genotypes, four norovirus GII genotypes, eight adenovirus genotypes, two astrovirus genotypes, and two sapovirus genotypes were detected. CONCLUSIONS: The Allplex assay showed high agreement with Seeplex and genotyping results, and was able to additionally detect sapoviruses. The Allplex assay could be useful in identifying viral gastrointestinal infections in patients with acute gastroenteritis symptoms.

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.

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.

Human Norovirus Genogroups Detected from Acute Gastroenteritis Patients in Seoul from May 2013 to April 2015

Norovirus is an important cause of acute nonbacterial gastroenteritis in communities worldwide. It was evaluated the prevalence of norovirus infections in patients with acute gastroenteritis occurring in Seoul from May 2013 to April 2015, with regular surveillance. 7.3% (252/3,485) of the fecal specimens were determined to be positive for noroviruses by reverse transcription-polymerase chain reaction (RT-PCR). Norovirus genogroup distribution was 19.1% (48/252) genogroup GI, 71.4% (180/252) genogroup GII, and 9.5% (24/252) genogroup G1+GII respectively. It was most norovirus detection rates from November 2013 to March 2015. And it was rotavirus 0.2% (7/3,485), astrovirus 0.03% (1/3,485), sapovirus 0.03% (1/3,485) and, it was non-detective on adenovirus. Norovirus genotypes identified were nine kinds of genogroup GI (GI-1, GI-2, GI-3, GI-4, GI-6, GI-7, GI-8, GI-12, GI-14) and eight kinds of genogroup GII (GII-2, GII-3, GII-4, GII-5, GII-6, GII-7, GII-14, GII-16, GII-17). The genetic characteristics of norovirus and the epidemiological patterns of a viral pathogen from acute gastroenteritis patients may give potentially effective data for epidemiological studies in Seoul, Korea.

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...

Prevalence of Human Astrovirus in Patients with Acute Gastroenteritis

The prevalence of human astroviruses was tested in patients with acute gastroenteritis by using conventional duplex reverse transcription (RT)-PCR and electrophoresis. Diarrheal fecal samples were collected from 9,597 patients at local hospitals in Seoul. The prevalence of astroviruses was 1.0% (94/9,597 patients; mostly infants), and that of sapoviruses was 0.1% (14/9,597 patients). Age- and gender-wise analyses were carried out on 29 astrovirus-positive patients having complete information on file regarding their age, gender, and other particulars. The results were higher in patients of ages 0 to 14 yr, and 69.0% of the astrovirus-positive patients were females, of which 69.2% were infants (0 to 12 months), and 61.5% were 1-4 yr old. Notably, in the case of 5 to 78-yr-old acute gastroenteritis patients, 100% were females.

Comparison among Known Severity Scoring Scales in the Evaluation of Acute Gastroenteritis in Children / 소아감염

PURPOSE: This study aimed to evaluate the disease severity of children suffering from gastroenteritis using different scales. The results are compared and subsequently classified on the basis of the type of virus causing the disease in order to investigate the differences in clinical characteristics and disease severity according to pathogen. METHOD: This study was conducted prospectively with patients under 5 years of age diagnosed with acute gastroenteritis and hospitalized at 9 medical institutions in 8 regions across the Republic of Korea. Disease severity was evaluated using the Vesikari Scale, the Clark Scale, and the modified Flores Scale. Fecal samples collected from patients were used to detect rotavirus and enteric adenovirus by enzyme immunoassay, and for RT-PCR of norovirus, astrovirus, and sapovirus. RESULTS: There were a total of 214 patients with a male : female ratio of 1.58 : 1, of which 35 were under the age of 6 months (16.4%), 105 were aged 6-23 months (49.1%), and 74 were aged 24-59 months (34.5%). The rate of concordance between the Vesikari and Clark Scales was 0.521 (P<0.001) and, in severe cases, the Vesikari Scale was 60.7% and Clark Scale was 2.3%, indicating that the Clark Scale was stricter in the evaluation of severe cases. CONCLUSIONS: In children with gastroenteritis, there were differences in disease severity based on the scale used. Therefore, to achieve consistent results among researchers, either only a single scale or a measure of all scales should be used to determine disease severity.

Clinical Features and Role of Viral Isolates from Stool Samples of Intussuception in Children / 대한소아소화기영양학회지

PURPOSE: To detect major acute gastroenteritis virus (rotavirus, norovirus, astrovirus, and enteric adenovirus) and non-enteric type of adenovirus (AdV) in the stools of intussusception patients and to investigate the clinical role of detected viruses. METHODS: From March 2012 to February 2013, major acute gastroenteritis virus and non-enteric type of AdV were isolated from stool samples that collected from 44 patients treated for intussusception in Chungnam National University Hospital. Patients were divided according to age and isolated virus. RESULTS: Virus was detected in 28 (63%) stool specimens. The virus detection rate was significantly lower in patients aged under 12 months (p = 0.04). Twenty-two patients (78.6%) had non-enteric adenovirus, 4 (14.3%) had norovirus, 1 (3.6%) had sapovirus, and 1 (3.6%) had astrovirus. AdV subgroup C (AdV 1, 2, 5, and 6) comprised the majority with 20 cases (90.9%). A monthly increment-and-decrement pattern of intussusception was similar to that of viral detection in the stool samples. Enema reductions were successful in 39 patients and surgical manual reductions were performed in 5 patients. Virus was detected in 24 patients (61.5%) of enema reduction group and 4 patients (80.0%) of surgical manual reduction group. All of the detected viruses were non-enteric adenovirus subgroup C (AdV 1, 5, and 6) in surgical reduction patients. CONCLUSIONS: The virus detection rate was high in the stools of intussusception patients. The pattern of seasonal intussusception occurrence rate was parallel with seasonal these viral detection rate in the stool samples. These findings suggest that viral infection plays an important role in the development of intussusception and further research is warranted.

Epidemiological features on 3 important viral diarrhea among children in Zhuhai during winter and spring / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the epidemiological characteristics of viral diarrhea of norovirus (NV), sapovirus (SV) and astrovirus (AstV) among children in Zhuhai during winter and spring.</p><p><b>METHODS</b>Stool specimens were collected from children with viral diarrhea in Maternal and Child Health Hospital of Zhuhai from November 21, 2009 to April 3, 2010. Nucleic acid of NV, SV and AstV from negative specimens of rotavirus and adenovirus were detected by using Reverse transcription-polymerase chain reaction (RT-PCR), and the types of positive samples of NV were also classified at the same time.</p><p><b>RESULTS</b>The total detection rate of the three viruses is 21.49 percent, the highest detection rate is 29.05% in December 2009, the lowest detection rate is 12.20% in February 2010, 87.96% of positive specimens were from children patients aged from 0 to 30 months. The season detection rate of NV, SV and AstV are 14.70%, 2.75% and 4.04% respectively. There were significant differences of NV and SV detection rates in every month of the season, whereas the AstV detection rate was comparatively stable. The highest detection rate of NV is 34.09% in children patients aged from 12 to 18 months, the highest SV detection rate is 12.5% in children patients aged from 60 to 120 months, and the highest AstV detection rate is 16.67% in children patients aged from 24 to 30 months. All the NV were belong to G II genogroup.</p><p><b>CONCLUSIONS</b>NV is one of the main pathogens causing viral diarrhea among children in Zhuhai during winter and spring, SV and AstV are also important pathogens. So we should strengthen the monitoring of viral diarrhea caused by NV, SV and AstV in infants and young children.</p>

HAND system-based four multiplex RT-PCR for simultaneous detection of four diarrhea viruses / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.</p><p><b>METHODS</b>Primers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.</p><p><b>RESULTS</b>The 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.</p><p><b>CONCLUSION</b>The established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.</p>

Prevalence of sapovirus infection among infant and adult patients with acute gastroenteritis in Tehran, Iran

This study investigated the prevalence of sapovirus infections in patient with acute gastroenteritis in Tehran, Iran. Sapovirus, a member of the family Caliciviridae is one of the major causative agents of viral gastroenteritis affecting both children and adult individuals. There isn't enough data about prevalence and genotypes of sapovirus infection in Tehran, the capital city of Iran. A total of 42 fecal samples were collected from patients with acute gastroenteritis from May to July 2009. RT nested- PCR was performed for screening. To genotype the sapovirus isolates, some positive samples were subjected to phylogenetic analysis by sequencing of fragments of viral capsid gene region. Sapovirus was detected in 5 of 42 stool specimens from patients with acute gastroenteritis. Saporvirus detected in this study was clustered into only one distinct genogroup I/2. Sapovirus GI/2 was predominant. Our results show that among the studied viruses responsible for this disease, sapovirus was a major viral isolate virus

Detection and genotype analysis of sapovirus associated with sporadic diarrhea in Shenzhen in 2009 / 南方医科大学学报

<p><b>OBJECTIVE</b>To conduct an epidemiological and genotype analysis of sapovirus (SaV) associated with sporadic diarrhea in Shenzhen in the year 2009.</p><p><b>METHODS</b>A total of 852 fecal samples were collected from sporadic cases of diarrhea in Shenzhen in 2009 and detected by reverse-transcription polymerase chain reaction (RT-PCR) using the primers of SLV5317/5749. The PCR products were analyzed with 1.5% agarose gel electrophoresis and sequenced to construct the phylogenetic tree.</p><p><b>RESULTS</b>Sixteen samples were found positive for SaV, with a positivity rate of 1.88%. Sequence analysis identified 8 isolates as SaV GI genotype (including 3 SaV GI.1 and 5 SaV GI.2), 7 as SaV GIV genotype, and 1 as GII genotype.</p><p><b>CONCLUSIONS</b>SaV infection is present in Shenzhen with GI as the predominant genotype. This is the first report of SaV GIV strains in China, which differs from the strains of Anhui-A141 and Beijing-CHN99/BJ360, suggesting the genotypic variety of SaV infection in China.</p>

Genetic diversity of porcine sapoviruses from Lulong county in China / 病毒学报

Porcine sapoviruses (SaVs), which belong to the family Caliciviridae, have been considered potential zoonotic agents for human infection, and several cases have been reported in Asian countries. In this study, a total of 200 porcine fecal samples collected from Lulong county of China were tested. Among 200 samples, porcine sapoviruses were detected by RT-PCR in 17 samples (8.5%) showing their circulation in China. 14 out of 17 positive sapovirus strains were genetically related to the genogroup III (GIII) and were further divided into three different clusters or genotypes according to the phylogenetic analysis. In addition, the remaining three sapovirus strains belonged to GVII (one strain) and a potential novel genogroup (two strains) according to the phylogenetic analysis and the nucleotide identity and amino acid identity. These data suggested the genetic diversity of porcine sapoviruses in China.

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.

Analysis of epidemiologic feature and genetic sequence of Sapovirus in China / 病毒学报

To investigate epidemiologic feature and genetic variance of Sapovirus among children in China, fecal specimens were collected from children under 5 years old with acute diarrhea from Feb 2006 to Jan 2007 in nine provinces including Anhui, Fujian et al. A total of 1,110 fecal samples were detected for Sapovirus by reverse transcriptase-PCR (RT-PCR). Ten samples (0.9%) were positive for Sapovirus. The PCR products were then sequenced and analysed by phylogenetic tree. The results indicated that the detected Sapovirus strains were classified into two genogroups and three genotypes, including G I/1, G I/3, G II/3.

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.