The Outer Capsid Protein VP4 of Murine Rotavirus Strain Eb Represents a Tentative New P Type
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In 1983, we isolated a porcine rotavirus (strain YM) that was prevalent in several regions of Mexico, as judged by the frequency of its characteristic electropherotype. By a focus reduction neutralization test, rotavirus YM was clearly distinguished from prototype rotavirus strains belonging to serotypes 1 (Wa), 2 (S2), 3 (SA11), 4 (ST3), 5 (OSU), and 6 (NCDV). Minor, one-way cross-neutralization (1 to 5%) was observed when antisera to the various rotavirus strains were incubated with rotavirus YM. In addition, the YM virus was not neutralized by neutralizing monoclonal antibodies with specificity to serotypes 1, 2, 3, and 5. The subgroup of the virus was determined to be I by enzyme-linked immunosorbent assay. To characterize the serotype-specific glycoprotein of the virus at the molecular level, we cloned and sequenced the gene coding for VP7. Comparison of the deduced amino acid sequence with reported homologous sequences from human and animal rotavirus strains belonging to six different serotypes further supported the distinct immunological identity of the YM VP7 protein.
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Abstract 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 positive for the group A rotavirus antigen detected by this ELISA. The rotavirus‐positive specimens were analysed with monoclonal antibodies specific for rotavirus VP6 subgroup I and II, and for VP7 serotypes G1–G4, G8, and G9. Of the rotavirus strains that could be subgrouped, VP6 subgroup I and II strains circulated at similar levels. Amongst the strains that could be serotyped, VP7 G9 strains predominated occurring in 17 cases, with G3 (n = 10) and G1 (n = 9) strains occurring in lower numbers. Four G8 strains were detected and only one G2 and no G4 strains were identified. This report extends the description of the global distribution of G9 rotavirus strains. J. Med. Virol. 67:608–612, 2002. © 2002 Wiley‐Liss, Inc.
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Molecular Epidemiology
Acute gastroenteritis
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A rotavirus-like virus (RVLV) was isolated from a diarrheic pig from an Ohio swine herd. This virus infected villous enterocytes throughout the small intestine of gnotobiotic pigs and induced an acute, transitory diarrhea. Complete virions were rarely observed in the intestinal contents of infected animals; the predominant particle detected by immune electron microscopy was a corelike particle 52 nm in diameter. The genome of the porcine RVLV was composed of 11 discrete segments of double-stranded RNA that produced an electropherotype distinct from the genome electropherotypes of reovirus, rotavirus, and porcine pararotavirus. Porcine RVLV was antigenically unrelated to rotavirus, porcine pararotavirus, or reovirus but was antigenically related to a bovine RVLV.
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ABSTRACT Sera from three separate healthy population cohorts were used to determine the incidence of group C rotavirus infections in 1,356 South Africans. Using an enzyme-linked immunosorbent assay based on a recombinant group C rotavirus VP6 protein, the total percent positivity was found to be 34.4% (range, 33 to 38%), with almost half of the population infected after the age of 20 years.
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Rotavirus Infections
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A series of reassortants was isolated from coinfection of cell cultures with a wild-type animal rotavirus and a "noncultivatable" human rotavirus. Wild-type bovine rotavirus (UK strain) was reassorted with human rotavirus strains D, DS-1, and P; wild-type rhesus rotavirus was reassorted with human rotavirus strains D and DS-1. The D, DS-1, and P strains represent human rotavirus serotypes 1, 2, and 3, respectively. Monospecific antiserum (to bovine rotavirus, NCDV strain) or a set of monoclonal antibodies to the major outer capsid neutralization glycoprotein, VP7 (of the rhesus rotavirus), was used to select for reassortants with human rotavirus neutralization specificity. This selection technique yielded many reassortants which received only the gene segment coding for the major neutralization protein from the human rotavirus parent, whereas the remaining genes were derived from the animal rotavirus parent. Single human rotavirus gene substitution reassortants of this sort represent potential live vaccine strains.
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Bovine rotavirus NCDV and simian rotavirus SA-11 represent two distinct rotavirus serotypes. A genetic approach was used to determine which viral gene segments segregated with serotype-specific viral neutralization. There were 16 reassortant rotaviruses derived by coinfection of MA-104 cells in vitro with the SA-11 and NCDV strains. The parental origin of reassortant rotavirus double-stranded RNA segments was determined by gene segment mobility in polyacrylamide gels and by hybridization with radioactively labeled parental viral transcripts. We found that two rotavirus gene segments found previously to code for outer capsid proteins vp3 and vp7 cosegregated with virus neutralization specificities.
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A four-layer solid phase enzyme-immunoassay (EIA) with antisera against Nebraska calf diarrhoea virus (NCDV) as immunoreagents was developed to detect human rotavirus antigens from stool specimens of patients with acute rotavirus gastroenteritis. Polystyrene beads were used as the solid phase, guinea-pig and rabbit anti-NCDV immunoglobulin as the catching and secondary antibody, and peroxidase-conjugated swine anti-rabbit immunoglobulin as the indicator antibody. A comparison of the developed NCDV-EIA with an identical EIA, using antisera against human rotavirus (HRV-EIA) instead of NCDV antisera, was made with 216 stool specimens positive or negative for rotavirus. A complete agreement was obtained between the two methods provided that appropriate confirmatory tests were included. The developed NCDV-EIA was as sensitive and specific for rotavirus as the HRV-EIA, and it allowed the detection of both established rotavirus types 1 and 2 from stools with equal sensitivity. The difficulties in cultivating human rotavirus in vitro for immunisation and the relative ease of growing NCDV in widely-used continuous cell lines make NCDV a good alternative in the preparation of the highly specific and sensitive rotavirus antisera required in immunoassays, and facilitate the setting-up methods for the routine diagnosis of rotavirus gastroenteritis by EIA or RIA in diagnostic virus laboratories.
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We analyzed the prevalence of rotavirus in 296 children age between 3 and 36 months who were hospitalized in 1994 with severe gastro-enteritis at two health centres for diarrhoea treatment in León, Nicaragua. Enteric viruses were detected in 96 (32.4%) of the children and rotaviruses were the most common pathogens detected in 84 (28%). The majority of rotavirus infections occurred in children less than 1 year old and all strains isolated belonged to subgroup II and had 'long' RNA patterns. Molecular epidemiology of 55 rotavirus strains revealed that all had the same RNA migration pattern and serotyping of 37 strains by PCR technology revealed that all isolates belonged to serotype 3. A significant observation was that only one electropherotype of rotavirus circulated. No non-group A rotaviruses were found by RNA gel electrophoresis. Adenoviruses were found by ELISA in 14 of 265 (5%) children and were most frequently detected during the 1st year of life. Of 103 faecal samples analyzed by electron microscopy, four contained small round structured viruses.
Molecular Epidemiology
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Twenty-two rotavirus isolates were recovered from piglets suffering from diarrhea. The isolates readily propagated in MA-104 cell cultures where they induced typical cytopathic effects (CPE) and possessed the physicochemicals properties of the members of rotavirus genus, Family Reoviridae. The electron microscopy study, conducted in MA-104 infected cells, revealed virus particles which possessed the peculiar morphology of rotavirus. The isolates were neutralized by a reference porcine rotavirus antiserum: all were devoid of hemagglutinating activity for red cells of swine, cattle, sheep, rabbit, chicken and guinea pig.
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