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first isolation of rotavirus associated with neonatal diarrhoea in guanacos (lama guanicoe) in the argentinean patagonia region.group a rotavirus (rv) and coronavirus (cv) are common viral pathogens associated with neonatal diarrhoea in numerous animal species. the purpose of this work was to investigate the presence of these viral agents in two farm populations of captured guanacos (lama guanicoe) in the argentinean patagonia region, that developed severe diarrhoea outbreaks. stool and serum samples were analysed for rv and bovine cv antigen and antibody enzyme-linked immunosorbent assay. rotavirus was detected in faece ...200111765807
nucleotide sequence analysis of rotavirus gene 11 from two tissue culture-adapted atcc strains, rrv and wa.we report here nucleotide sequence and characterization of gene 11 from two tissue culture-adapted atcc rhesus (rrv) and human (wa) strains of rotavirus. gene 11 sequence encodes a nonstructural protein, nsp5 and also encodes nsp6, from an out of phase open reading frame. sequence of rrv(atcc) gene 11 represents the first report from a rhesus rotavirus which has more than 90% homology at the nucleotide and deduced amino acid sequence level with that of its closely related simian sa11 strain. the ...200111778700
[evaluation of immunochromatography based rapid detection kit of rotavirus and adenovirus].we investigated the usefulness of rapidtesta rota-adeno (daiichi pure chemicals co., ltd., japan) for rapid detection of group a rotavirus and adenovirus simultaneously using immunochromatography with clinical samples. in investigation of the reaction of the kit to 5 strains of group a rotavirus, 13 strains of adenovirus and other intestinal viruses, specific lines were formed in red to group a rotavirus and blue to adenovirus. no cross reaction was observed with other intestinal virus. in measu ...200111806138
[a fusion protein of rotavirus vp6 and cholera toxin b subunit: expression in escherichia coli and analysis of biological activities].rotavirus infection is a major cause of dehydrating diarrhea in infants worldwide. the non-toxic cholera toxin b subunit(ctb), known as an immunomodulatory carrier, might help to stimulate mucosal immune response when coupled to rotavirus antigens in oral immunization. here we report for the first time the construction of a translational fusion of ctb gene 5' to the vp6 gene of a human rotavirus a(field strain t114), and expression of the ctb-vp6 fusion protein in e. coli bl21(de3). the expresse ...200111910752
[study on the genetic variations of the major neutralization antigen vp7 of group a rotavirus type g1 in china].to study the genetic variations of the major neutralization antigen vp7 of group a rotavirus, type g1 in china.200111986704
[etiological study on human rotavirus infections in children with acute gastroenteritis].to understand the etiology of human rotavirus infections in children with acute gastroenteritis in china.200112526305
[investigation of group a rotavirus infection in several cities in china and prevalence of vp7 serotype].to study the prevalence of group a rotavirus infection in china.200112526310
electropherotypes of rotaviruses in children less than 5 years old in abidjan, cote d'ivoire in 1997.rotaviruses are associated with gastroenteritis in human infants and the young of many species. in this study, we analysed the circulating strains of human rotavirus isolated from young children in abidjan by electrophoresis of the viral rna genome in agarose gels. rotavirus strains were identified in 33 children less than two years of age in the yopougon district of abidjan, cote d'lvoire. viral rna was extracted from the stools by phenol-chloroform treatment at 56 degrees c, followed by centri ...200117650045
nested reverse transcriptase-polymerase chain reaction for the detection of group a rotaviruses.rotaviruses are important pathogens associated with diarrhoeal diseases in almost all species of mammals. in the present study, a nested reverse transcriptase-polymerase chain reaction (rt-pcr) for the detection of group a rotaviruses was developed, which is based on a target region in gene segment 6. rotavirus strains of human, bovine, porcine, canine, feline, equine, and ovine origin were examined. furthermore several faecal specimens, in which rotavirus had already been detected using other m ...200212002423
a rapid method to produce high yields of purified rotavirus particles.a rapid purification method of rotavirus particles to high yield retaining the double shelled structure of infectious virus is described. group a rotavirus (uk strain) was concentrated through a cushion of colloidal silica (rho=1.10 g/cm(3)) or by precipitating with polyethylene glycol 8000. after concentration, infectious rotavirus was cleared from host cell proteins by density equilibrium centrifugation in gradients of colloidal silica using near vertical rotors. characterisation of purified v ...200212020788
rotavirus g serotypes and p[],g genotypes identified in cases of reinfection among children participating in a trial with rhesus-human reassortant tetravalent vaccine (rrv-tv) in belém, brazil.group a rotaviruses are the most important agents of severe diarrhea in children and infants worldwide. the aim of present study was to identify rotavirus g serotypes and p[],g genotypes in cases of reinfection among children who participated in a vaccine trial with the tetravalent rhesus-human reassortant rotavirus vaccine (rrv-tv 4 x 10(4) pfu/dose) in belém, brazil. from july 1990 to june 1992, 540 children received, at their first, third and fifth months of life, oral doses of either vaccine ...200212022436
molecular characterization of vp6 genes of human rotavirus isolates: correlation of genogroups with subgroups and evidence of independent segregation.a reverse transcription-pcr (rt-pcr) was established to amplify a 379-bp cdna fragment (nucleotides 747 to 1126, coding for amino acids 241 to 367) of the vp6 gene of group a rotaviruses associated with subgroup (sg) specificity. thirty-eight human rotavirus strains characterized with sg-specific monoclonal antibodies were subjected to vp6-specific rt-pcr, and pcr amplicons were used for sequencing. nucleic acid sequencing and phylogenetic analysis of the vp6 amplicons revealed two clusters, or ...200212050372
[outbreak of human rotavirus infection in an adult community].group a rotaviruses are known as the major cause of severe dehydrating diarrhoea in infants.200212073545
detection and genotyping of human group a rotaviruses by oligonucleotide microarray hybridization.a rapid and reliable method for the identification of five clinically relevant g genotypes (g1 to g4 and g9) of human rotaviruses based on oligonucleotide microarray hybridization has been developed. the genotype-specific oligonucleotides immobilized on the surface of glass slides were selected to bind to the multiple target regions within the vp7 gene that are highly conserved among individual rotavirus genotypes. rotavirus cdna was amplified in a pcr with primers common to all group a rotaviru ...200212089254
production of immunogenic vp6 protein of bovine group a rotavirus in transgenic potato plants.we report here the production of transgenic potato plants expressing the major capsid protein vp6 of bovine group a rotavirus (gar). transgenic plants under the control of a cauliflower mosaic virus 35s promoter, or a modified promoter linked to the tobacco mosaic virus 5'-untranslated sequence were positive for gar antigens by elisa. the expressed protein was consistent in size with vp6 of gar by western blot assay. the presence of the vp6 gene and its transcript was detected by pcr and rt-pcr. ...200212111435
circulation of the novel g9 and g8 rotavirus strains in nigeria in 1998/1999.an epidemiological survey investigating the prevalence of rotavirus infection in infants and young children with acute diarrhoea was undertaken in jos state, nigeria, between january 1998 and april 1999. in total, 672 faecal specimens were collected from children aged between 1 and 60 months with acute infantile gastroenteritis. the 10-20% stool suspensions were examined by an elisa for the presence of group a rotavirus antigen (rotavirus ideia, dako, uk). only 116 specimens (17.3%) were positiv ...200212116012
detection and quantitation of group a rotaviruses by competitive and real-time reverse transcription-polymerase chain reaction.a competitive reverse transcription-polymerase chain reaction (rt-pcr) was developed to detect and to quantitate the rna of group a rotaviruses. in the assay, a 433 bp fragment is amplified by a one-tube rt-pcr protocol using primers with binding sites located in a highly conserved region of segment 6 of the rotavirus genome. an in vitro synthesized rna with a 43-base deletion with respect to the wild-type sequence of this fragment was used as an internal control. using these transcripts as temp ...200212270660
molecular and immunological methods to detect rotavirus in formalin-fixed tissue.in 1999, a tetravalent rhesus-based rotavirus vaccine was withdrawn from the market after reports of intussusception cases among vaccinated infants. methods to detect rotavirus in formalin-fixed pathology specimens from such patients will be important in examining the possible associations between the vaccine and intussusception, in investigating fatalities caused by natural rotavirus infection, and in furthering our understanding of the pathogenesis of rotavirus disease. three different methods ...200212270663
causative pathogens of severe diarrhea in children.to investigate the enteropathogens in children with diarrhea attending salmaniya medical complex, bahrain.200212370713
virus diversity in a winter epidemic of acute diarrhea in france.in france, an epidemic peak of acute diarrhea is observed each winter. previous results suggested a viral etiology for these winter epidemics. we investigated the role of enteric viruses in acute diarrhea and their molecular diversity. one hundred sixty-one patients with acute diarrhea and 45 healthy patients (controls) from the general population were given a standardized questionnaire between december 1998 and may 1999. stool specimens were screened for group a and c rotaviruses, human caliciv ...200212409408
clinical utility of nested multiplex rt-pcr for group f adenovirus, rotavirus and norwalk-like viruses in acute viral gastroenteritis in children and adults.the diagnosis of viral gastroenteritis can be carried out by non-molecular techniques such as electron microscopy (em), enzyme-immunoassay and latex agglutination tests and various molecular techniques. normally molecular detection requires the use of three separate protocols to detect the three main causes of viral gastroenteritis, adenoviruses, rotaviruses and norwalk-like viruses (nlv) which have different types of nucleic acid. the development of a sensitive and specific assay which could de ...200212423697
detection and characterization of rotaviruses in hospitalized neonates in blantyre, malawi.in five separate fecal collections spanning three years, group a rotaviruses were detected by enzyme-linked immunosorbent assay in 35 (25%) of 142 specimens obtained from nondiarrheic, hospitalized neonates in blantyre, malawi. molecular characterization of each strain identified, for the first time in neonates, a short electropherotype, genotype p[6], g8 strain type, similar to the dominant, cocirculating community strain detected in symptomatic infants in blantyre. partial sequence analysis of ...200211923390
genetic analysis of group a rotaviruses: evidence for interspecies transmission of rotavirus genes.rotaviruses are the major cause of severe gastroenteritis in young children and animals. the rotavirus genome is composed of eleven segments of double-stranded rna and can undergo genetic reassortment during mixed infections, leading to progeny viruses with novel or atypical phenotypes. there are numerous descriptions of rotavirus strains isolated from human and animals that share genetic and antigenic features of viruses from heterologous species. in many cases, genetic analysis by hybridizatio ...200211928984
changes in the prevalence of rotavirus g and p types in diarrheic calves from the kagoshima prefecture in japan.g8 bovine group a rotavirus was the most predominant serotype in calf diarrheal fecal specimens examined between 1995 and 1996 in japan [vet. microbiol. 66 (1999) 301]. to date, no evidence that g8 was the most predominant in the typeable specimens has been observed outside japan. to investigate whether g8 continues to be as common as g6 and g10, the incidence of the main serotypes was determined in the same area (kagoshima prefecture) between 1997 and 1998 by reverse transcription-polymerase ch ...200211955784
rna-binding activity of the rotavirus phosphoprotein nsp5 includes affinity for double-stranded rna.phosphoprotein nsp5 is a component of replication intermediates that catalyze the synthesis of the segmented double-stranded rna (dsrna) rotavirus genome. to study the role of the protein in viral replication, his-tagged nsp5 was expressed in bacteria and purified by affinity chromatography. in vitro phosphorylation assays showed that nsp5 alone contains minimal autokinase activity but undergoes hyperphosphorylation when combined with the ntpase and helix-destabilizing protein nsp2. hence, nsp2 ...200211967345
a predominant role for norwalk-like viruses as agents of epidemic gastroenteritis in maryland nursing homes for the elderly.stool specimens from 156 maryland nursing home residents, who became ill during 20 outbreaks of gastroenteritis from november 1987 through february 1988, were analyzed. all tested negative for astroviruses, enteroviruses, group a rotaviruses, sapporo-like caliciviruses, and enteric bacteria (i.e., salmonella, clostridium, and shigella species). eighty-two (52%) were positive for norwalk-like viruses (nlvs), members of the family caliciviridae. six distinct genetic clusters within genogroups i an ...200211807686
human group c rotavirus: completion of the genome sequence and gene coding assignments of a non-cultivatable rotavirus.genome segments 1 and 2 of human group c rotavirus 'bristol' strain were sequenced and their gene-protein coding properties assigned. this work completed the genome sequence of a human group c rotavirus (17,910 bp) and allowed the full gene-protein coding assignment of the 11 segments of dsrna. gene 1 is 3309 bp in size and contains a single orf of 3272 nucleotides, encoding a protein of 1090 amino acids in length with a predicted molecular mass of 125 kda. comparison of the translated sequence ...200211864750
rotavirus nsp5: mapping phosphorylation sites and kinase activation and viroplasm localization domains.rotavirus nsp5 is a nonstructural protein that localizes in cytoplasmic viroplasms of infected cells. nsp5 interacts with nsp2 and undergoes a complex posttranslational hyperphosphorylation, generating species with reduced polyacrylamide gel electrophoresis mobility. this process has been suggested to be due in part to autophosphorylation. we developed an in vitro phosphorylation assay using as a substrate an in vitro-translated nsp5 deletion mutant that was phosphorylated by extracts from ma104 ...200211884570
genomic diversity of group a rotavirus strains infecting humans in eastern india.between 1998 and 2000, a total of 266 samples were found positive for group a rotaviruses by rna electrophoresis. samples were collected from patients admitted to two leading hospitals in calcutta. serotyping could be done only with 22% of the positive samples, leaving 78% untypeable. the g (vp7 genotypes) and p (vp4 genotypes) types were determined for 159 samples by reverse transcription and multiplex pcr. the predominant genotype was g1p[8] (20%), followed by g2p[4] (15%) and g4p[8] (6%). a n ...200211773108
group a rotavirus infection and age-dependent diarrheal disease in rats: a new animal model to study the pathophysiology of rotavirus infection.group a rotaviruses are major pathogens causing acute gastroenteritis in children and animals. to determine if group a rotavirus replicates and induces disease in rats, antibody-negative lewis neonatal or adult rats were inoculated orally with tissue culture-adapted human (wa, wi61, and hal1166), simian (rhesus rotavirus [rrv] and sa11), bovine (wc3), lapine (ala), or porcine (osu) rotavirus strains, wild-type murine (ec(wt)) rotavirus strain, or phosphate-buffered saline (pbs). rotavirus infect ...200211739670
epidemiological features of rotavirus infection among hospitalized children with gastroenteristis in ho chi minh city, vietnam.an epidemiological study of the g serotype and p genotype distribution of group a rotaviruses by using elisa and/or rt-pcr was conducted in children (aged 1 month to 15 years) with diarrhea that were admitted to the general children's hospital no. 1, ho chi minh city, vietnam from december 1999 to november 2000. the results showed that rotavirus is associated with 65.6% (889/1355) of diarrheal admissions. rotavirus infection mostly affected children under 2 years of age with a peak incidence in ...200312601768
distribution of human group a rotavirus vp7 and vp4 types circulating in seoul, korea between 1998 and 2000.three hundred forty-eight fecal specimens collected from young children with acute diarrhea in seoul, korea between january 1998 and february 2000 were examined for g and p types. of these, 205 samples (59%) were confirmed as group a rotavirus by elisa for the detection of vp6 antigen. confirmed rotavirus isolates were characterized using g serotyping elisa and rt-pcr methodologies for g and p genotyping of the outer capsid proteins vp7 and vp4, respectively. serotyping of the outer capsid prote ...200312696125
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%) faecal specimens positive for rotavirus a, most of them collected from hospitalized children. a sig ...200312700858
serotyping of human group a rotaviruses in nara prefecture, japan. 200312711829
genetic characterization of a novel, naturally occurring recombinant human g6p[6] rotavirus.a binary classification system has been established for group a rotaviruses, with the viral capsid protein vp7 defining g types and vp4 defining p types. at least 15 g types and 21 p types have been isolated globally with various g and p combinations. most of the currently circulating human rotaviruses belong to g1p[8], g2p[4], g3p[8], and g4p[8]. we report a human rotavirus strain (b1711) with a novel genotypic vp7/vp4 combination of g6p[6]. this unique rotavirus was isolated from a 13-month-ol ...200312734253
detection of a mammalian-like group a rotavirus in diarrhoeic chicken.the present investigation describes detection of a mammalian-like electropherogroup a rotavirus in chicken with diarrhoea. this also records the first detection of a rotavirus in an avian species from india. during the investigation 75 diarrhoeic faecal samples collected from adult chicken were screened for the presence of group a rotavirus antigen by sandwich elisa. all three samples positive for rotavirus antigen revealed 11 bands of rna in polyacrylamide gel electrophoresis (page). in contras ...200312742711
emergence of novel human group a rotavirus g12 strains in india.three rare human g12 strains were detected from diarrheic clinical samples of children (<8 months of age) in calcutta during a routine surveillance study of rotaviral diarrhea in india. the vp7 genes of g12 strains and their products showed maximum homology (97 to 99% at the nucleotide level and 98% at the amino acid level, respectively) with those of two recently reported g12 strains (from the united states and thailand) but lesser homology with those of prototype g12 strain l26.200312791925
construction and characterization of rhesus monkey rotavirus (mmu18006)- or bovine rotavirus (uk)-based serotype g5, g8, g9 or g10 single vp7 gene substitution reassortant candidate vaccines.group a rotaviruses are the single most important etiologic agents of severe diarrhea of infants and young children worldwide and have been estimated to be responsible for approximately 650,000-800,000 deaths annually in children <5-year-old in the developing countries. thus, the development of a safe and effective rotavirus vaccine has been a global public health goal. epidemiologic surveillance of rotavirus vp7 (g) serotypes-genotypes conducted in various populations throughout the world has r ...200312798644
a human vaccine strain of lamb rotavirus (chinese) nsp4 gene: complete nucleotide sequence and phylogenetic analyses.a lamb strain of rotavirus has recently been licensed for use in china as a live vaccine to prevent rotavirus diarrhea in children. as rotavirus nsp4, especially the cytotoxic domain alone is considered to be associated with diarrhea, we sequenced gene segment 10, which encodes nsp4, of lamb rotavirus. comparative analyses was performed to identify differences from human rotavirus strains, that might be associated with attenuation, and to ascertain whether the lamb rotavirus gene fits among the ...200312803470
group a rotavirus in sewage samples from barcelona and cairo: emergence of unusual genotypes.the presence of rotavirus strains in sewage samples from cairo, egypt (november 1998 to october 1999), and barcelona, spain (november 1998 to december 2002), was investigated by using a generic molecular detection method based on amplification of a vp6 gene fragment. overall, 85.7 and 66.9% of the sewage samples from cairo and barcelona, respectively, were positive. positive samples were characterized further, and vp7 and vp4 genotypes were determined. although 30% of the positive samples from c ...200312839761
detailed computational analysis of a comprehensive set of group a rotavirus nsp4 proteins.rotavirus infection causes diarrhea to humans, animals and birds. the nsp4 protein of group a rotavirus has been recognized as a viral enterotoxin. this single protein plays important roles in viral pathogenesis and morphogenesis. domains involved in structure and biologic functions have been proposed mainly based on the sa11 strain, a prototype of group a rotavirus. nsp4 has been classified into different genotypes based on sequence homology. these analyses are based on representative strains s ...200312876455
clinical manifestations of the rotavirus infection and his relation with the electropherotypes and serotypes detected during 1998 and 1999 in merida, yucatan, mexico.group a rotavirus (rv) is associated with acute infectious diarrhea (aid) in children and adults. the clinical manifestations of rv infection are classified as slight, moderate and severe and could be the results of differing rotaviral serotypes. attempts have been made to correlate the severity of the infection with specific rv groups, subgroups (sg) serotypes and electropherotypes, but the results have been inconclusive.200312878087
molecular and serological characterization of group a rotavirus isolates obtained from hospitalized children in goiânia, brazil, 1998-2000.fecal samples positive for rotavirus group a ( n=120) were analyzed by enzyme immunoassay-monoclonal antibody (eia-mab) serotyping and/or reverse transcription/multiplex polymerase chain reaction (pcr) amplification to determine the prevalence of the [p] and g genotypes. the most prevalent g genotype/serotype detected was g1 (76.7%), followed by g2 (5.0%). six samples were characterized as g9 by multiplex pcr, and one sample was characterized as g3 by eia-mab. the combinations of [p] and g genot ...200312884075
identification of a bisegmented double-stranded rna virus (picobirnavirus) in calf faeces.to determine the incidence of rotavirus infection among dairy herds in the state of são paulo, brazil, 576 faecal samples obtained from calves aged 1-45 days with and without diarrhoea, reared on 63 dairy cattle farms, were analyzed. polyacrylamide gel electrophoresis (page) identified 28 samples positive for group a rotavirus, while four samples, two diarrhoeic and two non-diarrhoeic, showed a bisegmented genome with a typical picobirnavirus pattern. electron microscopy revealed spherical virus ...200312902184
serological survey of anti-group a rotavirus igm in uk adults.rotaviral associated disease of infants in the uk is seasonal and infection in adults not uncommon but the relationship between these has been little explored. adult sera collected monthly for one year from routine hospital samples were screened for the presence of anti-group a rotavirus immunoglobulin m class antibodies as a marker of recent infection. anti-rotavirus igm was seen in all age groups throughout the year with little obvious seasonal variation in the distribution of antibody levels. ...200312948372
relationship between group a porcine rotavirus and management practices in swine herds in ontario.a case-control epidemiological study was conducted to determine whether an increased diagnostic rate for group a rotavirus in swine herds in ontario was associated with specific management factors. the number of new herds tested per year and the proportion of new positive herds increased between 1994 and 1997. herd size was larger and weaning age was younger in rotavirus-positive herds compared with rotavirus-negative herds. pigs raised in all-in all-out nurseries were 3.4 times more likely to h ...200313677596
rotavirus detection and characterisation in outbreaks of gastroenteritis in aged-care facilities.although rotavirus is a major cause of gastroenteritis in children, its role in adult gastroenteritis and the sensitivity of different methods for its detection in specimens collected from adults are less well understood.200314522072
an outbreak of diarrhoea in one-week-old piglets caused by group a rotavirus genotypes p[7],g3 and p[7],g5.thirty-two group a isolates of rotavirus detected in faecal samples from diarrhoeic piglets, were selected for p and g genotyping using a multiplex rt-pcr. ten isolates, from animals less than 8 days old, characterized an outbreak of diarrhoea caused by group a rotavirus in animals. p[7],g3 (crw8-like) and p[7],g5 (osu-like) genotypes were detected in 5 animals each. isolates of a group a rotavirus of genotypes compatible with the osu prototype were those most frequently identified in single inf ...200314582749
p genotypic identification of human group a rotaviruses. 200314583647
a novel type of vp4 carried by a porcine rotavirus strain.the gene encoding the vp8* trypsin-cleavage product of the vp4 protein of porcine rotavirus strain a34 was sequenced, and the predicted amino acid (aa) sequence was compared to the homologous region of all known p genotypes. the aa sequence of the vp8* of strain a34 shared low identity, ranging from 39% (bovine strain b223, p8[11]) to 76% (human strain 69m, p4[10]), with the homologous sequences of representative strains of the remaining 21 p genotypes. phylogenetic relationships showed that the ...200314585340
microarrays for genotyping human group a rotavirus by multiplex capture and type-specific primer extension.human group a rotavirus (hrv) is the major cause of severe gastroenteritis in infants worldwide. hrv shares the feature of a high degree of genetic diversity with many other rna viruses, and therefore, genotyping of this organism is more complicated than genotyping of more stable dna viruses. we describe a novel microarray-based method that allows high-throughput genotyping of rna viruses with a high degree of polymorphism by multiplex capture and type-specific extension on microarrays. denature ...200314605152
isolation and characterization of dually reactive strains of group a rotavirus from hospitalized children.seven rotavirus strains dually reactive to serotype g1- and g2-specific monoclonal antibodies (mabs) from hospitalized children with rotavirus diarrhea were culture adapted. six strains were neutralized with g1 antiserum to a higher titer than that of g2, and one was neutralized with g1 and g2 antisera to equal titers. of these, four strains were also neutralized with g6 antiserum. five strains with short rna pattern could not be serotyped, and the remaining two strains with long rna pattern wer ...200314605180
evaluation of a human group a rotavirus assay for on-site detection of bovine rotavirus.neonatal diarrhea induced by bovine group a rotavirus causes significant economic loss in the dairy and beef industry due to increased morbidity and mortality, treatment costs, and reduced growth rates. the objective of this study was to evaluate a human group a rotavirus assay (immunocardstat rotavirus [ics-rv]) as an on-site diagnostic test for bovine rotavirus. when used with a collection of bovine diarrhea samples submitted to the virology section of the diagnostic center for population and ...200312517863
detection and characterization of human rotavirus among children with diarrhoea in botswana.this study reports the detection, for the first time, of human rotavirus in stools of children and the molecular characterization of isolated circulating strains in botswana. we collected 249 stool samples between 1999 and 2001 from children with diarrhoea in three health districts of botswana and examined them for the presence of rotavirus antigens and particles. group a rotavirus antigen was detected in 43 of 249 (17%) of the samples tested by enzyme-linked immunosorbent assay. of the 43 child ...200314641850
acute diarrhea in paraguayan children population: detection of rotavirus electropherotypes.group a rotavirus infections were detected in 93 of 410 fecal samples from children with acute diarrhea, admitted in three main hospitals of asunción, paraguay, from august 1998 to august 2000. most of the rotavirus-infected patients were admitted during the winter season in the three epidemic years. the rotavirus infection rate was highest in infants from 6 to 23 months of age. in the 93 samples examined, 10 different rotavirus electropherotypes were recognized, but two of them largely predomin ...200314658840
molecular analysis of the vp7, vp4, vp6, nsp4, and nsp5/6 genes of a buffalo rotavirus strain: identification of the rare p[3] rhesus rotavirus-like vp4 gene allele.we report the detection and molecular characterization of a rotavirus strain, 10733, isolated from the feces of a buffalo calf affected with diarrhea in italy. strain 10733 was classified as a p[3] rotavirus, as the vp8* trypsin cleavage product of the vp4 protein revealed a high amino acid identity (96.2%) with that of rhesus rotavirus strain rrv (p5b[3]), used as the recipient virus in the human-simian reassortant vaccine. analysis of the vp7 gene product revealed that strain 10733 possessed g ...200314662959
risk factors for norovirus, sapporo-like virus, and group a rotavirus gastroenteritis.viral pathogens are the most common causes of gastroenteritis in the community. to identify modes of transmission and opportunities for prevention, a case-control study was conducted and risk factors for gastroenteritis attributable to norovirus (nv), sapporo-like virus (slv), and rotavirus were studied. for nv gastroenteritis, having a household member with gastroenteritis, contact with a person with gastroenteritis outside the household, and poor food-handling hygiene were associated with illn ...200314720397
rotavirus associated acute diarrhoea in hospitalized children in dibrugarh, north-east india.faecal specimens from 202 children below 5 years with acute diarrhoea hospitalized in assam medical college from april, 1999 to march, 2000 were examined in regional medical research centre (icmr), dibrugarh to know the prevalence of rotavirus diarrhoea and molecular pattern of viral strains from different localities of dibrugarh using double antibody sandwich elisa and sds-page analysis. human group a rotaviruses were detected in 47 (23.27%) specimens and 33 of 41 (80.49%) positive specimens we ...200315022939
human group c rotaviruses identified in kenya.rotavirus is associated with acute infantile gastro-enteritis in infants and young children worldwide. rotavirus is associated with the high cases of morbidity and mortality and it is estimated that up to 650,000 deaths in young children occur annually in the less developed countries. approximately a quarter of these deaths occur in african children yet minimal data exist on the circulating rotavirus strains in africa.200316167719
rotavirus - a retrospective study of incidence at the hospital universiti sains malaysia (husm).the present study, analyzes data from 1991 to 2000 for rotavirus infection among children with diarrhoea and acute gastroenteritis admitted to the hospital universiti sains malaysia (husm). the latex slide agglutination test was used for the detection of rotavirus antigens. out of 1097 stool samples tested, 207 samples or 18.8 % were found to be positive for rotavirus. the infection occurred most frequently in infants and young children from 6 months to 2 years of age. the infection was recorded ...200323386803
sequence analysis of the guanylyltransferase (vp3) of group a rotaviruses.the rna segment encoding the guanylyltransferase (vp3) from 12 group a rotavirus isolates has been sequenced following rt-pcr and molecular cloning of the full-length amplicons produced. alignment of the derived amino acid sequences including those of the four vp3 sequences available from genbank revealed two levels of sequence divergence. virus isolates from humans showed greater than 94% sequence identity, whereas those isolated from different mammalian species showed as low as 79% sequence id ...200415039535
quantitation of group a rotavirus by real-time reverse-transcription-polymerase chain reaction: correlation with clinical severity in children in south india.the epidemiology and pathogenesis of rotaviruses are not completely understood, although recent developments in polymerase chain reaction (pcr) techniques now make it possible to quantify the viral load during an infective episode and investigate its relevance to clinical features of the disease. we studied rotavirus-positive stool samples collected from 10 children without symptoms of gastroenteritis and from 81 children with acute gastroenteritis and in whom the clinical severity of disease wa ...200415042658
molecular epidemiology of viral gastroenteritis in asia.acute gastroenteritis is one of the most common diseases in humans, and continues to be a significant cause of mortality and morbidity worldwide. recently the estimates of mortality associated with diarrhea declined, however the majority of deaths still occur in developing countries and thus urgent intervention is needed for the prevention of these diseases. in asian countries it is very important to study the distribution, transmission and characteristics of prevalent viruses in order to produc ...200415056259
g and p genotypes of group a rotavirus strains circulating in calves in brazil, 1996-1999.fifty fecal samples from calves with diarrhea, positive for group a rotavirus by enzyme immunoassay (eia) and polyacrylamide gel electrophoresis (page) were analyzed by a nested multiplex reverse transcription/polymerase chain reaction (nested multiplex/rt-pcr) for identification of p and g genotypes. samples were collected between 1996 and 1999 from eight dairy and/or beef cattle herds located in the mato grosso do sul, são paulo, goiás and paraná states, brazil. complete genotyping was possibl ...200415066719
[rotavirus a infections and reinfections: genotyping and vaccine implications].to identify rotavirus a and the most prevalent g and p genotypes in children with acute diarrhea, and to the describe the occurrence of rotavirus infection and reinfection.200415079181
g and p genotypes of group a rotavirus from diarrhoeic calves born to cows vaccinated against the ncdv (p[1],g6) rotavirus strain. 200415107035
homotypic and heterotypic serum isotype-specific antibody responses to rotavirus nonstructural protein 4 and viral protein (vp) 4, vp6, and vp7 in infants who received selected live oral rotavirus vaccines.homotypic and heterotypic serum isotype-specific antibody responses to rotavirus enterotoxin nonstructural protein (nsp)-4, independent neutralization antigens viral protein (vp)-4 and vp7, and group a rotavirus common antigen vp6 were analyzed by an immunocytochemistry assay in infants who received 1 of several live oral rotavirus vaccines. significant serum immunoglobulin (ig) a and igg antibody responses to homotypic and/or heterotypic nsp4s of genotype [a], [b], or [c] were detected after va ...200415122520
molecular genotyping of irish rotavirus strains.rotavirus is the primary etiological agent of gastroenteritis in infants and young children worldwide. in developing countries, it is estimated that rotavirus is responsible for one-third of all diarrhea-associated hospitalizations and 873,000 deaths annually. in industrialized countries, where mortality from rotavirus is low, infection is widespread, and nearly all children experience an episode of rotavirus diarrhea in the first 3-5 yr of life. rotaviruses have important antigenic specificitie ...200415156021
an outbreak of gastroenteritis during school trip caused by serotype g2 group a rotavirus.between may14 and 18, 2001, there was an outbreak of acute gastroenteritis involving 45 school children out of a total of 107 (aged 11-12 years) attending a 3-day school trip. the epidemic curve characterized by a rapid onset and decline with a single peak incidence over a 5-day period resembled the pattern typical of a food-borne gastroenteritis outbreak. epidemiological and virological investigations concluded, however, that this outbreak was caused by a single strain of serotype g2 group a ro ...200415170643
rotavirus serotype g9 strains belonging to vp7 gene phylogenetic sequence lineage 1 may be more suitable for serotype g9 vaccine candidates than those belonging to lineage 2 or 3.a safe and effective group a rotavirus vaccine that could prevent severe diarrhea or ameliorate its symptoms in infants and young children is urgently needed in both developing and developed countries. rotavirus vp7 serotypes g1, g2, g3, and g4 have been well established to be of epidemiologic importance worldwide. recently, serotype g9 has emerged as the fifth globally common type of rotavirus of clinical importance. sequence analysis of the vp7 gene of various g9 isolates has demonstrated the ...200415220453
verification of sensitivity and specificity of group a rotavirus detection in piglets faeces with monoclonal blocking elisa methods.monoclonal antibodies to group a rotavirus vp6 protein were prepared and used for verification of three blocking enzyme-linked immunosorbent assay (elisa) modifications to detect rotavirus a. selected competitive blocking elisa (cb-elisa) and electron microscopy (em) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. rotavirus was detected in 43 samples (22.2%) by cb-elisa method, whereas in 26 (13.4%) samples by em examination. however, of 26 samples posit ...200415228549
human infection with a p[14], g3 lapine rotavirus.group a rotaviruses are the main cause of severe diarrhoea in humans and animals throughout the world. we report the first description of a clinically apparent infection with a p[14], g3 rotavirus (strain b4106) in a hospitalized 6-year-old child. the vp7 gene of the b4106 strain had the closest sequence similarity (94% and 97% on the nucleotide and amino acid level, respectively) with strain 30/96 (p[14], g3), a lapine rotavirus isolated in an italian rabbit in 1996 while the vp4 gene had the c ...200415231381
g and p genotypes of rotavirus circulating among children with diarrhea in the colombian northern coast.a study on the prevalence of rotavirus g and p genotypes was carried out based on 253 stool specimens obtained from children living in the colombia northern coast region who were less than 3-years-old and who suffered from acute diarrhea. a previous study had detected the presence of rotavirus a in 90 (36.5%) of the 246 samples tested by enzyme immunoassay (eia), and these strains were investigated in the present study. of these, 50 strains yielded an rna electropherotype, most of which (80.0%) ...200415248159
virus diversity and an outbreak of group c rotavirus among infants and children with diarrhea in maizuru city, japan during 2002-2003.a total of 236 fecal specimens collected from infants and children with gastroenteritis in maizuru city, japan from july 2002 to june 2003, were tested for the presence of rotaviruses, noroviruses, sapoviruses, astroviruses, and adenoviruses by rt-pcr, page, rpha, and latex agglutination methods. among diarrheal viruses detected, group a rotavirus was the most prevalent (32.2%; 76 of 236) followed by norovirus gii (21.2%; 50 of 236), group c rotavirus (10.2%; 24 of 236), adenovirus (3.8%; 9 of 2 ...200415258985
group b rotaviruses similar to strain cal-1, have been circulating in western india since 1993.generally, group a rotaviruses are the most common cause of paediatric diarrhoea. however, group b rotavirus, adult diarrhoea rotavirus (adrv), was found to be involved in epidemics of severe gastroenteritis in several areas of china during 1982-1983 and had resulted in more than one million cases among adults as well as older children. human group b rotavirus has been rarely reported outside china, but has been detected first from five adults with diarrhoea in kolkata, india during 1997-1998 (s ...200415310177
application of a molecular method for the detection of group a rotaviruses in raw and treated water.group a human rotaviruses (hrvs) are the most important aetiological agents of acute viral gastroenteritis in infants and young children in both developing and industrialised countries. rotaviruses are resistant to many chemical disinfectants and reportedly survive well in treated tapwater and sewage. in this study a group a specific reverse transcriptase-polymerase chain reaction (rt-pcr) followed by a nested-pcr was applied for the detection of hrvs in raw and treated drinking-water samples dr ...200415318513
molecular characterization of novel p[14],g8 bovine group a rotavirus, sun9, isolated in japan.in this study, a novel bovine group a rotavirus (rv-a), sun9, isolated from calf diarrhea in the tochigi prefecture, japan, was characterized genetically by the sequence analysis of the genome segments encoding vp4 and vp7. the nucleotide and deduced amino acid sequences of the genome segments encoding vp4 and vp7 of sun9 revealed high homology with p[14] human and lapine rv-as (80.2-88.7% and 90.9-94.8%) and g8 bovine and human rv-as (83.1-95.5% and 92.3-98.2%). sun9 was also classified into p[ ...200415325086
development of rt-multiplex pcr assay for detection of adenovirus and group a and c rotaviruses in diarrheal fecal specimens from children in china.rotavirus, adenovirus, norovirus, sapovirus and astrovirus are considered to be significant global enteropathogens associated with sporadic cases and outbreaks of acute gastroenteritis. therefore, a rapid and sensitive assay is preferred to screen for the presence of these viruses in diarrheal fecal specimens. in a previous study, we developed a reverse transcription single-round multiplex polymerase chain reaction (rt-smpcr) assay for the simultaneous detection of norovirus (genogroup i, genogr ...200415478645
effects of intrabodies specific for rotavirus nsp5 during the virus replicative cycle.intracellular antibodies or intrabodies (icabs) have great potential in protein knockout strategies for intracellular antigens. in this study, they have been used to investigate the role of the rotavirus non-structural protein nsp5 in the virus replication cycle. intracellular antibody-capture technology was used to select single-chain fv format (scfv) icabs against an nsp5 mutant. five different specific icabs were selected and expressed in ma104 cells, in the scfv format, as cytoplasmic- and n ...200415483242
rotavirus strain surveillance in latin america: a review of the last nine years.latin america will likely be the first area in the developing world where rotavirus vaccine will be introduced into the routine childhood immunization schedule. in anticipation of that goal, we reviewed the distribution of group a rotavirus genotypes in latin america to understand the diversity of strains to be targeted by vaccines and to identify novel strains that may pose challenges for vaccines.200415502697
the vp6 protein of rotavirus interacts with a large fraction of human naive b cells via surface immunoglobulins.immunity to human group a rotavirus (rv), a major cause of viral gastroenteritis in infants, involves b lymphocytes that provide rv-specific antibodies. additionally, some arguments suggest that naive b cells could be implicated in the first steps of the immune response against rv. the aim of our study was to analyze the interaction of vp6 and vp7 rv capsid proteins with human b cells depending on the immune status of the individual, i.e., naive or rv experienced. for this purpose, a two-color v ...200415507636
[molecular epidemiologic survey of rotaviruses from infants and children with diarrhea in shanghai].to investigate molecular epidemiologic features of rotaviruses circulating in shanghai, china.200414990097
diarrhea caused by rotavirus in children less than 5 years of age in hanoi, vietnam.group a rotaviruses are the major cause of diarrhea in young children worldwide. from march 2001 to april 2002, 836 children less than 5 years of age were investigated in hanoi, vietnam. this included 587 children with diarrhea and 249 age-matched controls. group a rotavirus was identified in 46.7% of the children with diarrhea and 3.6% of the controls, which was a significant difference. within the diarrhea group, the highest prevalence was seen in children from 13 to 24 months of age, and the ...200415583308
genetic and serological characterization of novel serotype g8 bovine group a rotavirus strains isolated in japan.g8 bovine group a rotaviruses isolated in japan were genetically and serologically characterized. the vp7 gene nucleotide and amino acid sequences revealed high identity with each other. all japanese g8 strains were classified into the same lineage in the phylogenetic analysis based on vp7 gene sequences. antisera to four japanese g8 strains neutralized other g8 strains, but their neutralizing titers were between 8-fold lower and 2-fold higher than homologous strains. these results suggest that ...200415585957
an elisa optimized for porcine epidemic diarrhoea virus detection in faeces.monoclonal antibodies to porcine epidemic diarrhoea virus (pedv) membrane protein m were prepared and used for the comparative assessment of three blocking elisa variants to detect pedv. the competitive blocking elisa (cb-elisa) format showed the highest sensitivity, allowing detection of 10(2.5) plaque-forming units of pedv/ml in culture medium. its specificity was verified by inclusion of control samples containing transmissible gastroenteritis virus (tgev) and rotavirus a in each analysis. ei ...200415607079
genetic variability of human rotavirus strains isolated from eastern and northern india.an epidemiological study was conducted in eastern and northern india to determine the genomic diversity of rotaviruses in these parts of the country. in 2001, a total of 126 group a rotavirus positive samples were detected from children below 4 years of age with diarrhoea from kolkata, dibrugarh and bhubaneswar in eastern india, and chandigarh, a city in northern india. all the samples were genotyped for vp7 (g-type) and vp4 (p-type) gene by reverse transcription (rt) and multiplex pcr using dif ...200414635025
accumulation of rotavirus vp6 protein in chloroplasts of transplastomic tobacco is limited by protein stability.rotavirus vp6 is a highly immunogenic major capsid protein that may be useful as a subunit vaccine. the expression of a bovine group a rotavirus vp6 cdna was examined in tobacco chloroplasts following particle bombardment. constructs containing the vp6 cdna under the control of plastid rrn or psba promoters, or the escherichia coli trc promoter, were inserted, together with the aada selectable marker gene, between the rbcl and accd genes of the tobacco plastid genome. the 40-kda vp6 protein accu ...200417147617
determination of bovine rotavirus g genotypes in kashmir, india.rotavirus ribonucleic acid (rna) was extracted from ten faecal samples of diarrhoeic calves positive for group a rotavirus by enzyme-linked immunosorbent assay (elisa). a portion of the extracted rna was run in polyacrylamide gel to determine the presence of rotaviral rna and the rest subjected to reverse transcriptase-polymerase chain reaction (rt-pcr) to generate the near full length vp7 gene. only six samples yielded the desired product. the amplified products were subjected to g-typing by pc ...200415861888
rotavirus rna electropherotype in different states in malaysia for the year 2000 and 2001.a total of 157 stool samples were examined for group a rotaviruses in diarrheic children admitted to 8 different major hospitals in malaysia. the overall incidence rate in this study was 19.7% (31 of 157) with a variation of 9.5% to 39.1% in different locations. majority of the infections detected were in those under 2 years of age and there were fewer admissions in the older age group. the stool samples were initially screened for rotavirus group a by latex agglutination method and followed by ...200415559163
rotavirus infection among infants and young children in chennai, south india.group a rotavirus has been recognized as the major etiologic agent of childhood gastroenteritis in infants and young children worldwide. rapid progress towards the development of an efficacious rotavirus vaccine has warranted extensive epidemiological studies on rotavirus serotypes that cause severe disease in developing and developed countries and to monitor the emergence of newer and unusual strains in different geographical settings that could represent variants not covered by existing vaccin ...200417642741
a rapid and accurate assay for assessing the cytotoxicity of viral proteins.a fluorescence-based assay is presented for measuring the cytoxicity of viral proteins added exogenously to cells. the assay is based on the use of two fluorescent dyes, calcein-am and ethidium homodimer (etd-1) to specifically stain living and dead cells respectively and employs fluorescence activated cells sorting (facs) to achieve a rapid and accurate measurement of the cytotoxic capacity of a potential viral toxin. the assay has been developed using the group b homologue (adrv-nsp4) of the n ...200515899525
dominating prevalence of p[8],g1 and p[8],g9 rotavirus strains among children admitted to hospital between 2000 and 2003 in budapest, hungary.group a rotaviruses are the main cause of acute dehydrating diarrhea in children, responsible for high mortality in developing countries and a significant socio-economic burden associated with treating the disease in developed countries. two rotavirus vaccine candidates predicated on either homotypic or heterotypic protection have undergone clinical trials recently and await licensure for routine use. in anticipation of a future vaccination campaign in hungary, the diversity of rotaviruses colle ...200515902709
design and selection of an intrabody library produced de-novo for the non-structural protein nsp5 of rotavirus.intracellular antibodies or intrabodies have great potential in protein knockout strategies for intracellular antigens. we applied the intracellular antibody capture technology for the direct selection in yeast of a mouse scfv library (v(l)-v(h) format) constructed from animals immunised with recombinant non-structural protein nsp5 of rotavirus. we selected five different intracellular antibodies (icabs), which specifically recognize delta2, an nsp5 deletion mutant used as bait. the anti-nsp5 ic ...200515907924
sequence analysis of the vp7 and vp4 genes identifies a novel vp7 gene allele of porcine rotaviruses, sharing a common evolutionary origin with human g2 rotaviruses.during an epidemiological survey encompassing several porcine herds in saragoza, spain, the vp7 and vp4 of a rotavirus-positive sample, 34461-4, could not be predicted by using multiple sets of g- and p-type-specific primers. sequence analysis of the vp7 gene revealed a low amino acid (aa) identity with those of well-established g serotypes, ranging between 58.33% and 88.88%, with the highest identity being to human g2 rotaviruses. analysis of the vp4 gene revealed a p[23] vp4 specificity, as it ...200515914225
identification of the novel lapine rotavirus genotype p[22] from an outbreak of enteritis in a hungarian rabbitry.application of improved molecular techniques in the detection and characterization of rotavirus strains has led to the recent description of several new combinations, specificities, and genetic variants of the outer capsid genes, vp7 and vp4. in spite of the enormous diversity of mammalian rotavirus strains, the few lapine rotaviruses characterized to date, appear to carry a narrow range of such antigen combinations; only p[14], g3 and, based on a more recent study, p[22], g3 rotaviruses have pr ...200515936106
oral immunization with pbsvp6-transgenic alfalfa protects mice against rotavirus infection.a critical factor in edible plant-derived vaccine development is adequate expression of the exogenous antigens in transgenic plants. we synthesized a codon-optimized gene (svp6) encoding the vp6 protein of human group a rotavirus and inserted it into the alfalfa genome using agrobacterium-mediated transformation. as much as 0.28% of the total soluble protein of the pbsvp6-transgenic alfalfa was svp6. female balb/c mice were gavaged weekly with 10 mg of transgenic alfalfa extract containing 24 mi ...200515992851
serological characterization of novel p11[14],g8 bovine group a rotavirus, sun9, isolated in japan.in this study, a novel bovine group a rotavirus (borv-a), sun9, isolated from calf diarrhea in tochigi prefecture, japan, was serologically characterized by a cross-neutralization assay, and serological surveillance by using its reassortant was performed on cattle bred in japan. the g serotype of sun9 was identified as g serotype 8 based on the one- or two-way serological relationships observed in sun9 and other g8 strains. the p serotype of sun9 was identified as p serotype 11 based on the one- ...200515996788
characterization of a novel p[25],g11 human group a rotavirus.a novel rotavirus strain (dhaka6) isolated from a 21-year-old bangladeshi male patient was characterized by sequence analysis of its vp7 and vp4 gene segments. phylogenetic analysis of the vp7 gene of the dhaka6 strain revealed a common evolutionary lineage with porcine g11 rotavirus strains. this isolate is the first reported g11 rotavirus strain infecting a human host. comparison of the vp4 gene sequences with all currently recognized 24 different p genotypes revealed only low nucleotide (54 t ...200516000437
outbreak of acute gastroenteritis associated with group a rotavirus and genogroup i sapovirus among adults in a mental health care facility in japan.an outbreak of acute gastroenteritis consisting of 57 cases occurred in a mental health care facility in takasaki city, japan during 6th february and 27th march 2002. a total of 18 fecal specimens collected from 17 residents and one member of the medical staff during this outbreak were tested for the presence of viral enteropathogens by rt-pcr and latex agglutination. group a rotavirus and sapovirus were detected in 5 out of 18 fecal specimens (55.6%). to our knowledge, this is the first finding ...200515648058
a porcine g9 rotavirus strain shares neutralization and vp7 phylogenetic sequence lineage 3 characteristics with contemporary human g9 rotavirus strains.of five globally important vp7 (g) serotypes (g1-4 and 9) of group a rotaviruses (the single most important etiologic agents of infantile diarrhea worldwide), g9 continues to attract considerable attention because of its unique natural history. serotype g9 rotavirus was isolated from a child with diarrhea first in the united states in 1983 and subsequently in japan in 1985. curiously, soon after their detection, g9 rotaviruses were not detected for about a decade in both countries and then reeme ...200515661150
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