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polyacrylamide gel electrophoresis of rna compared with polyclonal- and monoclonal-antibody-based enzyme immunoassays for rotavirus.polyacrylamide gel electrophoresis (page) of rotaviral rna, a sensitive and highly specific test for detecting rotavirus in stool, was compared with two commercially available enzyme immunoassays (eias), monoclonal (pathfinder) and polyclonal (rotazyme ii). stool samples from 204 children with nosocomial diarrhea were tested for rotavirus by both eias and by page of rna extracted from raw stools or 10% stool suspensions. samples which tested positive by either eia but were negative by page were ...19882830303
a combined enzyme immunoassay for rotavirus and adenovirus (eiara).a combined enzyme immunoassay for rotavirus and adenovirus (eiara) was developed as a double-antibody sandwich assay in which test samples are added to plastic wells coated with rotavirus or adenovirus goat antibody. the presence of antigens is detected by mixed-guinea pig antisera to the same viruses followed by rabbit anti-guinea pig igg conjugated with peroxidase and subsequently by ortho-phenylenediamine substrate. titrations of rotavirus (sa11) and adenovirus type 2 tissue culture grown vir ...19852982891
[procedures for enhancing the sensitivity of the counterimmunoelectro-osmophoresis method in diagnosing viral infections]. 19852998097
hemagglutination with ovine rotavirus. brief report.the virus was grown in ma 104 cells, a stable cell line derived from embryonic rhesus monkey kidney, and tested for hemagglutination (ha) with erythrocytes of a variety of species at 4 degrees c, room temperature and 37 degrees c. ha was observed at all temperatures with chicken, sheep, rabbit, guinea pig and human erythrocytes but not with horse, cattle, goat, swine and goose erythrocytes. ha reaction was inhibited by specific antiserum. some factors involved in the ha and ha-inhibition (hi) we ...19863015075
immune rabbit sera do not recognize antigenic variants of caprine arthritis-encephalitis virus isolated during persistent infection.isolates of caprine arthritis-encephalitis virus recovered 12 to 18 months after infection of goats were tested for neutralization by using rabbit antisera directed to the original inoculum virus. only slight differences in neutralization kinetics between the long-term isolates and the original virus were detected, indicating that heterologous rabbit antisera do not detect antigenic variation of the neutralizing epitopes of caprine arthritis-encephalitis virus.19836196294
avidin-biotin radioimmunoassay for human rotavirus.rias have a number of advantages which make them ideally suited for use in diagnostic microbiology. these advantages include sensitivity, objectivity, and versatility. however, the widespread application of rias has been limited by the instability of the reagents required for the performance of available solid-phase rias. the relatively short half-life of gamma-emitting isotopes is particularly a problem in cases where multiple antigens must be assayed, since distinct radioactively labeled reage ...19836313823
enzyme-linked immunosorbent assays based on polyclonal and monoclonal antibodies for rotavirus detection.we describe two enzyme-linked immunosorbent assays for rotavirus antigen in feces, which were designed to be as sensitive and specific as possible, and easy to use anywhere. both are indirect methods, using the antibody capture method, but the second assay utilizes a rotavirus group-specific monoclonal "detecting" antibody instead of the hyperimmune polyvalent guinea pig antisera used in the first assay. both tests were found to be more sensitive than electron microscopy for detecting virus. to ...19846321549
isolation and preliminary characterization of a caprine rotavirus.five cytopathic rotavirus strains were isolated in ma104 cells from stool specimens of kids with diarrhoea. pre-treatment of the virus with trypsin and incorporation of low levels of trypsin in the maintenance medium were important for the successful cultivation of the strains in these cells. the isolates were shown to be group a rotaviruses by antigenic reactivity with a group a monoclonal antibody. this was confirmed by the migration patterns of the viral rna genome during polyacrylamide gel e ...19947501360
rotavirus in saanen goats.rotavirus particles were detected in the stools of 8 out of 12 saanen kids with either diarrhoea or an increased rectal temperature. no other pathogenic organisms could be recovered from these kids. this is believed to be the first report of rotavirus infection in goats in southern africa.19947595922
prevalence of antibody to group b (atypical) rotavirus in humans and animals.enzyme-linked immunosorbent assays were developed for the detection of group b rotavirus antigen and antibody. the specificities of both assays were evaluated for antigens and serum specific for rotavirus groups a to d. serum collected in the united kingdom from different animal species exhibited the following high prevalence of group b rotavirus-specific antibody: pigs, 97%; cattle, 71%; sheep, 91%; and goats, 91%. in human serum, a lower prevalence of group b-specific antibody was detected; se ...19873029164
rotavirus infection in calves, piglets, lambs and goat kids in trinidad.faecal samples from diarrhoeic and non-diarrhoeic calves, piglets, lambs and goat kids were collected and screened by a latex agglutination test to detect the presence of group a rotavirus antigen. of a total of 470 animals screened, 138 (29.4%) had faecal samples positive for rotavirus antigen. the prevalences of infection were 27.7% (73/264) in calves, 27.8% (45/162) in piglets, 48.6% (18/37) in lambs and 28.6% (2/7) in goat kids. rotavirus antigen was not detected in calves and lambs < 1 week ...19948044669
isolation and identification of rotaviruses as aetiological agents of neonatal diarrhoea in kids. electrophoretical characterization by page.six strains of cytopathogenic enteric virus for the continuous cell line ma-104 were isolated from samples of the diarrhoeic stools of kids (capra hyrcus) with acute enteritis. the samples were obtained from two stock farms (rm and si) in central italy. four viruses from the rm stock were identified as rotaviruses of the antigenic group a by the elisa test performed with sa-11 antirotavirus monoclonal antibodies and the direct immunofluorescence test with igg bovine antirotavirus. electrophoreti ...19938396195
development of indigenous elisa for rotavirus diagnosis & its comparison with commercial kit.preparation of reagents for the diagnosis of rotavirus by enzyme linked immunosorbent assay (elisa) was carried out at the national institute of virology (niv), pune. this is a double antibody sandwich elisa test. the coating antibody was raised in guinea pig against sa-11 (simian rotavirus), and is used at 1:20,000 dilution. the indicator antibody was also raised against sa-11 in rabbits. the rabbit preimmune serum is used as negative control serum. both, pre- and post-immune rabbit antisera ar ...19938406647
an outbreak of diarrhoea associated with atypical rotaviruses in goat kids.the presence of rotaviruses was investigated in the faeces of 31 goat kids in a dairy herd that experienced an outbreak of severe diarrhoea which caused dehydration, anorexia and prostration in seven (22.6 per cent) of them. all the affected animals were two to three days old. a group a-specific elisa failed to detect rotaviruses in any of the samples but the characteristics electropherotype of group b rotaviruses was observed by electrophoresis in polyacrylamide gels in six of the animals. a hi ...19958525112
caprine and bovine b rotaviruses in western france: group identification by northern hybridization.in a survey of coronavirus and rotavirus-induced neonatal diarrhoea in 373 calves from 284 farms, nine (2.4%) of the animals were found to be infected with non-group a rotaviruses when their faeces were analysed in a commercial elisa and by page. seven out of eight 1-4- day-old kids from a single farm were also infected with similar viruses. in a mini-page, all the viruses displayed group b- and/or e-like 4223 electrophoretypes. in northern hybridizations with cdna chemiluminescent probes specif ...19968721296
role of enteric pathogens in the aetiology of neonatal diarrhoea in lambs and goat kids in spain.faeces samples from diarrhoeic and non-diarrhoeic lambs and goat kids aged 1-45 days were examined for enteric pathogens. cryptosporidium parvum was detected in both diarrhoeic lambs (45%) and goat kids (42%) but not in non-diarrhoeic animals. f5+ (k99+) and/or f41+ escherichia coli strains were isolated from 26% and 22% of the diarrhoeic lambs and goat kids, respectively, although these strains, which did not produce enterotoxins st i or lt i, were found with similar frequencies in non-diarrhoe ...19968760970
sequence analysis and in vitro expression of genes 6 and 11 of an ovine group b rotavirus isolate, kb63: evidence for a non-defective, c-terminally truncated nsp1 and a phosphorylated nsp5.an ovine group b rotavirus (gbr) isolate, kb63, was isolated from faeces of a young goat with diarrhoea in xinjiang, people's republic of china. sequence determination and comparison of genes 6 and 11 with the corresponding sequences of gbr strains adrv and idir showed that they were the cognate genes encoding nsp1 and nsp5, respectively. while the overall identities of nucleotide sequences between these two genes and the corresponding genes of strains adrv and idir were in the range 52.6-57.2%, ...199910466806
development of dot-elisa for the detection of human rotavirus antigen and comparison with rna-page.development of a simple, specific, rapid and inexpensive dot-elisa test for diagnosis of rotaviral antigen in stool samples.200111264748
isolation, serologic and molecular characterization of the first g3 caprine rotavirus.we isolated a rotavirus in cell culture, named the grv strain, from a stool specimen of a korean goat with diarrhea, and performed an in-depth characterization. at various passage levels in cell culture, the grv strain retained its pathogenicity for goat kids, thereby for the first time establishing that a caprine rotavirus can cause diarrhea in goat kids. the grv strain grew to a high titer and agglutinated group o human erythrocytes. the grv vp7 protein was 96% identical with the rrv (simian r ...200312664291
molecular characterization of the first isolation of rotavirus in guanacos (lama guanicoe).rotaviruses (rvs) are a major cause of neonatal diarrhoea in humans and numerous animal species. we report g-type, p-type and phylogenetic analysis of two rv strains isolated from newborn guanacos (lama guanicoe) with acute diarrhoea in argentina. isolates were group a g8 rv. phylogenetic analysis showed a close relationship to other g8 bovine rv from japan, u.s.a. and switzerland. p-types were p[1] and an unusual p[14], related to human and goat p[14] strains. this is the first report of a p[14 ...200415449134
specific labelling of human rotaviruses and adenoviruses with gold-igg complexes.human rotaviruses and adenoviruses were specifically labelled by an indirect immuno gold staining technique for electron microscopy. rabbit antisera against human rota- and adenoviruses were used as the primary antibody and goat anti-rabbit igg-gold complexes as secondary antibody. optimal conditions for the procedure were established by testing serial dilutions of the antisera. precoating of electron microscopy grids with bovine rotavirus antibody decreased nonspecific background staining and a ...19852416768
complete genome constellation of a caprine group a rotavirus strain reveals common evolution with ruminant and human rotavirus strains.this study reports the first complete genome sequence of a caprine group a rotavirus (gar) strain, go34. the vp7-vp4-vp6-vp1-vp2-vp3-nsp1-nsp2-nsp3-nsp4-nsp5 genes of strain go34, detected in bangladesh, were assigned to the g6-p[1]-i2-r2-c2-m2-a11-n2-t6-e2-h3 genotypes, respectively. strain go34 was closely related to the vp4, vp6-7 and nsp4-5 genes of bovine gars and the nsp1 gene of go34 to an ovine gar. strain go34 shared low nucleotide sequence identities (<90 %) with vp2-3 genes of other g ...201020505013
putative canine origin of rotavirus strain detected in a child with diarrhea, taiwan.abstract rotavirus g3p[3] strains have been reported from a variety of species including humans, cats, dogs, monkeys, goats, and cows. here, we report the characterization of the first human g3p[3] rotavirus from east asia identified in a 2-year-old child who was treated in a hospital's emergency ward in taiwan in february 2005. sequence and phylogenetic analysis demonstrated a close genetic relationship between the vp4, vp6, vp7, and nsp4 genes of taiwanese g3p[3] strain 04-94s51 and an itali ...201122022813
induction of castration by immunization of male dogs with recombinant gonadotropin-releasing hormone (gnrh)-canine distemper virus (cdv) t helper cell epitope p35.immunocastration is a considerable alternative to a surgical castration method especially in male animal species for alleviating unwanted male behaviors and characteristics. induction of high titer of antibody specific for gonadotropin-releasing hormone (gnrh) correlates with the regression of testes. fusion proteins composed of canine gnrh and t helper (th) cell epitope p35 originated from canine distemper virus (cdv) f protein and goat rotavirus vp6 protein were produced in e. coli. when these ...200515785119
detection of antibody to turkey coronavirus by antibody-capture enzyme-linked immunosorbent assay utilizing infectious bronchitis virus antigen.an antibody-capture enzyme-linked immunosorbent assay (elisa) for detection of antibody to turkey coronavirus (tcv) utilizing infectious bronchitis virus (ibv) antigen was developed. anti-tcv hyperimmune turkey serum and normal turkey serum were used as positive or negative control serum for optimization of the elisa system. goat anti-turkey immunoglobulin g (light plus heavy chains) conjugated with horseradish peroxidase was used as detector antibody. the performance of the elisa system was eva ...200011006996
characterization by polymerase chain reaction of ruminant rotaviruses isolated in italy.several group a rotaviruses isolated in italy from cattle, buffalos and goats were characterized by polymerase chain reaction assay for g- and p-type. g6 and p5 were the types most frequently recovered. the genotypes of buffalo and goat strains were similar to those of cattle isolates.199910322609
sensitive solid-phase immune electron microscopy double-antibody technique with gold-immunoglobulin g complexes for detecting rotavirus in cell culture and feces.a new solid-phase immune electron microscopy double-antibody colloidal-gold technique (spiemdagt) was developed and compared with direct electron microscopy, direct immune electron microscopy, and enzyme immunoassay for detecting rotavirus. guinea pig and rabbit antirotavirus antisera were used as capture and detector antibodies, respectively, and goat anti-rabbit immunoglobulin g-gold complexes were employed as a label. animal rotavirus in cell culture media and human virus in stool specimens w ...19902161868
rotavirus quantification: a comparison between the enumerative and the mpn techniques.a commonly used technique to isolate rotaviruses from a water environment involves the inoculation on cell culture associated with the revelation of the viral multiplication by indirect immunofluorescence. the results of the experiment using this methodology may be expressed either with an enumerative technique or with a mpn technique. we have found that the enumerative method provided the highest yield as opposed to the mpn method by a ratio close to 3.7. according to the experimental condition ...19957619199
are human p[14] rotavirus strains the result of interspecies transmissions from sheep or other ungulates that belong to the mammalian order artiodactyla?a limited number of human g6p[14] rotavirus strains that cause gastroenteritis in humans have been isolated in europe and australia. the complete genome sequences were determined for five of these human strains--b10925-97 (isolated in belgium in 1997), 111/05-27 (italy, 2005), pa169 (italy, 1987), mg6 (australia, 1993), and hun5 (hungary, 1997)--and their genetic relatedness to animal rotavirus strains was evaluated by sequencing the complete genome of the sheep rotavirus ovr762 (g8p[14]; spain, ...200919153225
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