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PubMed1999

An outbreak of group C rotavirus gastroenteritis among children attending a day-care centre in Belém, Brazil.

In August 1993, an outbreak of group C rotavirus-associated gastroenteritis occurred among children attending a day-care centre in Belém, Brazil. Of the 64 children, 21 (33%) became ill. Group C rotavirus was identified in faecal specimens from 8 (38%) children with diarrhoea by electron microscopy (EM) and an enzyme immunoassay (EIA), using antibodies specific to the Cowden strain of porcine group C rotavirus. By polyacrylamide gel electrophoresis (PAGE), a pattern similar to that of group C rotavirus was observed in 5 (62.5%) of the 8 EM- and EIA-positive samples. These 5 faecal samples were confirmed to be positive for group C rotavirus by reverse transcriptase-polymerase chain reaction, using specific VP6 and VP7 primers. This is the first report of an outbreak of diarrhoea in North Brazil associated with group C rotavirus. These findings suggest that group C rotavirus may be an important aetiological agent of diarrhoea in this region, which requires further study.

Brazil↗
PubMed2004

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, genogroup II), sapovirus and astrovirus in fecal specimens (Yan et aL, 2003). Recently, we developed another RT multiplex PCR for one-step amplification of all subgenera A to F adenoviruses, and group A and C rotaviruses. In this study, a total of 207 fecal specimens collected from children with acute gastroenteritis between December 2001 and April 2003 in Yunnan Province, China were examined for the presence of adenoviruses, and group A and C rotaviruses, by RT-multiplex PCR. The detection rate of these three viruses was 55.1% (114 out of 207 specimens), among which adenovirus and group A and C rotaviruses were identified in 11, 101 and 1 fecal specimen, respectively. Furthermore, one specimen was found to be positive for co-infection with adenovirus and group A rotavirus. An epidemic of acute gastroenteritis was also identified as peaking mainly in October and November. Taken together, our results clearly indicate that this novel assay provides a potentially rapid and convenient tool for epidemiologic investigation of diarrhea caused by adenovirus and group A and C rotaviruses.

Acute Disease↗
PubMed1996

Detection of group C rotavirus in infants with extrahepatic biliary atresia.

The purpose of this retrospective study was to examine liver tissue from patients with cholestatic disease for the presence of group C rotavirus RNA. The reverse transcriptase-polymerase chain reaction (PCR) for genes 5 and 6 was used, and the PCR products were subjected to liquid hybridization with a 32P-labeled probe. A second amplification with nested primers was also used. Samples from 32 subjects (20 with biliary atresia or choledochal cyst and 12 controls) were tested. Ten of 20 biliary atresia patients were positive for group C rotavirus RNA; no controls were positive (P < .003). Three of the positive patients were positive for both genes 5 and 6. Six of the 10 had > 1 sample that was positive. These data suggest a possible relationship between group C rotavirus and extrahepatic biliary atresia in the 10 patients in whom virus RNA was detected.

Autoradiography↗
PubMed1991

Molecular cloning, sequence analysis and coding assignment of the major inner capsid protein gene of human group C rotavirus.

The VP6 gene of human group C rotavirus was cloned and sequenced. Hybridization to the human group C and the porcine group C/Cowden dsRNA genomes assigned this coding sequence to segment 5. The complete human VP6 sequence contained an open reading frame of 1185 nucleotides (395 amino acids; deduced Mr 44,669 Da). The protein sequence demonstrated low homology with the group A VP6 sequences (41.7 to 42.7%) and high homology (88.9%) with the porcine group C VP6 sequence. However, the protein sequence alignments revealed a region of 10 amino acids that were significantly different between the human and the porcine group C viruses.

Amino Acid Sequence↗
PubMed2004

Seroepidemiology of group C rotavirus infection in England and Wales.

A total of 3199 serum samples collected in 1993--1994 from two population cohorts from England and Wales were tested for the prevalence of IgG antibodies specifically directed against group C rotavirus VP6. Seroprevalence was 39% (95% confidence intervals: 37.0-40.4%). Seroprevalence was highest (46.0%) in the oldest age group (61-70 years of age). Infection with group C rotaviruses occurred at an earlier age and with higher incidence in rural than in urban populations. These results may suggest transmission from animals to humans, however further work is required to identify the reservoir of group C rotavirus for human infection.

Adolescent↗
PubMed1991

Adaptation and serial passage of porcine group C rotavirus in ST-cells, an established diploid swine testicular cell line.

A porcine group C rotavirus (strain Cowden-AmC-1) was adapted and serially passaged in an established diploid swine testicular cell line (ST cells). Growth of group C rotavirus in ST cells, which in maintenance medium required trypsin, but not pancreatin resulted in cytopathic effect characterized by cellular stranding and subsequent cell lysis. Active replication and assembly were confirmed by RNA profile analysis, immune electron microscopy and immunofluorescence. Adaptation of non-group A rotavirus to a continuous cell line has not previously been reported and should facilitate progress in diagnostic procedures and vaccine development.

Adaptation, Physiological↗
PubMed1999

Sequence analysis of the gene encoding VP4 of a bovine group C rotavirus: molecular evidence for a new P genotype.

Nucleotide sequence of the bovine group C rotavirus Shintoku strain gene 3 was determined. Segment 3 is 2253 nucleotides (nt) in length and contains a long open reading frame (ORF) beginning at nt 22 and terminating at nt 2223. This ORF encodes a polypeptide of 733 amino acids with a predicted molecular mass of 83 kDa. The deduced gene 3 amino acid sequence shares 79% and 73% identities with VP4 of the porcine Cowden and human Bristol strains, respectively. Lack of high amino acid sequence homology in VP4 of bovine, porcine, and human group C rotaviruses indicates that the Shintoku strain represents a new P genotype.

Amino Acid Sequence↗
PubMed1992

Group C rotavirus requires sialic acid for erythrocyte and cell receptor binding.

Current immunological and biochemical information regarding the hemagglutinin and virus-cell interactions of rotavirus is obtained exclusively from studies with group A rotaviruses. In this study, I report that the immunologically and genetically distinct group C rotavirus also possesses a hemagglutinin. The viral hemagglutinin was identified on a cultivable porcine group C rotavirus strain (strain AmC-1) by using agglutinated human and guinea pig erythrocytes. Neuraminidase treatment of fresh human erythrocytes or blocking with glycophorin A or fetuin prevented hemagglutination. Infection of swine testicular cells with group C AmC-1 virus was also prevented by glycophorin A, fetuin, and neuraminidase treatment, suggesting that sialic acid constitutes an essential part of the cell receptor.

Animals↗
PubMed1990

Porcine group C rotavirus as a cause of neonatal diarrhea in a Quebec swine herd.

A porcine group C rotavirus was found to be the unique cause of a problem of enzootic neonatal diarrhea in a minimal disease herd composed of 190 sows on a continuous farrowing program. During the outbreaks of diarrhea, 10 to 80% of the litters were affected with a morbidity rate of 100% and case fatality rates of 5 to 10%. Clinical signs began 24 to 48 h after birth and were characterized by a profuse yellow diarrhea lasting a few days. Piglets from different outbreaks of diarrhea were necropsied. They had multifocal villous atrophy in the small intestine, especially in the ileum. Group C rotavirus was demonstrated by direct immunofluorescent staining of frozen intestinal sections and by polyacrylamide gel electrophoresis of viral RNA extracted from the intestinal contents of diarrheic piglets. The infection with clinical illness and lesions was reproduced experimentally in newborn piglets by oral inoculation of a suspension prepared from a pool of intestinal contents from diarrheic piglets.

Animals↗
PubMed1999

Seroepidemiology of human group C rotavirus in South Africa.

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.

Adolescent↗
PubMed2001

Detection of group C rotavirus in children with acute gastroenteritis in Blantyre, Malawi.

Among 606 children who were treated for acute gastroenteritis at the Queen Elizabeth Central Hospital in Blantyre, Malawi, Group C rotavirus (Gp C RV) was detected by enzyme-linked immunosorbent assay in fecal specimens from 16 (3.9%) of 408 inpatients and in 4 (2.0%) of 198 outpatients. Thirteen (65%) children excreting Gp C RV were coinfected with Group A rotavirus.

Acute Disease↗
PubMed1999

Comparisons of nucleotide and deduced amino acid sequences of NSP4 genes of virulent and attenuated pairs of group A and C rotaviruses.

The NSP4 protein of rotavirus is a nonstructural glycoprotein and has a crucial function in virus morphogenesis during infection of host cells. It was recently reported that NSP4 may also function as a viral enterotoxin in the induction of rotavirus diarrhea by causing Ca++ influx in the cytoplasm of the infected cells. We sequenced and analyzed two (Wa and M strains) pairs of NSP4 genes of virulent (v) and attenuated (a) (after 30 to 40 passages in cell culture) human group A rotaviruses and a pair of NSP4 genes of virulent and attenuated porcine group C rotavirus (Cowden strain). These strains were previously identified as virulent (induce diarrhea) or attenuated (no diarrhea) in a gnotobiotic pig model of rotavirus infection [Bohl et al. (4), Saif et al. (13), Ward et al. (17)]. The NSP4 genes of the Wa, M and Cowden strains were amplified with RT-PCR using a proof reading polymerase (Tli) and the RT-PCR product was sequenced directly. Analysis of the NSP4 deduced amino acid sequences showed that only 3 (Wa) and 2 (M and Cowden) amino acids differed between the virulent and attenuated strains. For the Wa strain, the changes from the virulent to attenuated strain were in amino acids 13 (V to A), 16 (L to S) and 34 (P to L); in the M strain, the difference was in amino acids 53 (T to I) and 104 (K to E), and in the Cowden strains, amino acids 50 (L to F) and 97 (D to N) differed between virulent and attenuated strains. To our knowledge, this is the first sequence comparison between NSP4 of a virulent and attenuated pair of group C rotaviruses. The potential impact of these few amino acid changes on the pathogenesis of the NSP4 protein for piglets is unclear, relative to previous findings in mice (1), but requires further study using purified recombinant NSP4 proteins or peptides.

Amino Acid Sequence↗
PubMed1999

Demonstration of group C rotaviruses in fecal samples of diarrheic dogs in Germany.

Three out of 9 fecal samples from diarrheic dogs, which were positive for rotavirus by electron microscopy, revealed RNA-migration patterns identical to those of porcine group C rotavirus when studied by polyacrylamide gel electrophoresis (PAGE). The results were further confirmed by solid phase immuno electron microscopy (SPIEM). This is the first report of the occurrence of group C rotaviruses in dogs.

Animals↗
PubMed1991

Detection of group B and C rotaviruses by polymerase chain reaction.

We adapted the polymerase chain reaction (PCR) to detect the noncultivatable group B and C rotaviruses and introduced a simple and convenient technique to purify viral RNA from stool specimens. Double-stranded RNA present in stool extracts was purified by adsorption to hydroxyapatite and was used as the template for reverse transcription and polymerase amplification. Primer pairs specific for group B (gene 8) and group C (gene 6) rotaviruses were selected to amplify group-characteristic sizes of cDNA copies readily identifiable in ethidium bromide-stained agarose gels. These primer pairs were used separately in individual PCR assays or were pooled with a primer pair specific for group A rotavirus (gene 9) in a combined PCR assay for the simultaneous detection of all three rotavirus groups. The method was very sensitive and was used to identify both human and porcine strains of group B and C rotaviruses in stool specimens. A second PCR amplification with internal group-specific primers served to increase further the sensitivity of the test and to confirm the diagnostic results obtained in the first amplification.

Base Sequence↗
PubMed2002

Detection of human group C rotaviruses in Nigeria and sequence analysis of their genes encoding VP4, VP6, and VP7 proteins.

In a survey on the etiology of acute gastroenteritis in infants and young children in Nigeria, group C human rotaviruses were detected in two of 112 rotavirus positive stool specimens collected between 1999 and 2000. The VP7, VP6, and VP4 genes of the two Nigerian human group C rotavirus strains (Jajeri and Moduganari) were sequenced in this study. Comparative sequence analysis with other published human group C rotaviruses showed that the genes encoding the three structural proteins were remarkably conserved in primary structure with few mutations. The VP4 and VP7 genes from the two Nigerian strains were related more closely to each other than to those of other published strains, and formed a separate cluster on the phylogenetic tree. In contrast, it was of note that VP6 gene of strain Moduganari was related more closely to the Brazilian strain Belem than to the other Nigerian strain Jajeri. This is the first report of identification of human group C rotavirus in Nigeria and constitutes the first sequence data of human group C rotaviruses in the African continent.

Antigens, Viral↗