scholarly journals Electropherotypes and G-Types of Group A Rotaviruses Detected in Children with Diarrhea in Lagos, Nigeria

ISRN Virology ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Christianah Idowu Ayolabi ◽  
David Ajiboye Ojo ◽  
George Enyimah Armah
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Migration Pattern ◽  
Genomic Diversity ◽  
Rt Pcr ◽  
Group A Rotaviruses ◽  
Group A ◽  
Stool Samples ◽  
Health Care Centers ◽  
Virus Isolates

Approximately over 500,000 children die annually due to severe dehydrating diarrhea caused by rotaviruses. This work investigated rotavirus infection among children less than 5 years with diarrhea in Lagos and determined the circulating electropherotypes and genotypes of the virus isolates. Three hundred and two (n=302) stool samples from children below 60 months were collected from different hospitals and health care centers in Lagos and subjected to enzyme immunoassay (EIA) to determine the presence of Group A rotavirus, RT-PCR to determine the G-types, and polyacrylamide gel electrophoresis (PAGE) to determine the electropherotypes. The results show that 60.3% of the samples showed distinct rotavirus RNA migration pattern, having long electropherotypes (55.3%) of seven variations dominating over the short electropherotypes (44.5%). Six different G-types were detected (G1, G2, G3, G4, G9, and G12). Serotypes G1 and G12 showed long electropherotypic pattern while G2, G3, and G9 exhibited either short or long electropherotype. All G4 detected show short electropherotypic pattern. In conclusion, information on the genomic diversity and RNA electropherotypes of rotaviruses detected in children with diarrhea in Lagos is reported in this study.

scholarly journals Rotavirus genotyping in gastroenteritis cases of an infantile population from Western Brazilian Amazonia

2012 ◽  
Vol 45 (4) ◽  
pp. 520-522 ◽  
Author(s):  
Maria Sandra Moura Costa ◽  
Paulo Afonso Nogueira ◽  
Gleicienne Félix Magalhães ◽  
Paula Taquita ◽  
Luis André Mariúba ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Brazilian Amazon ◽  
Rt Pcr ◽  
Rotavirus A ◽  
Positive Stool ◽  
Group A ◽  
Stool Samples

INTRODUCTION: During the period from 2000 to 2002, 79 rotavirus-positive stool samples were collected from children presenting diarrhea in the Western Brazilian Amazon. METHODS: Molecular characterization of the G and P genotypes was performed using RT-PCR and electropherotyping analysis by polyacrylamide gel electrophoresis. RESULTS: A total of 59 samples were confirmed as group A rotavirus. A long electrophoretic profile was exhibited by the G1P[8], G3P[8], and G4P[8] genotypes. The G1P[8] genotype was found in greater proportion. The short electropherotype was exhibited only by G2 genotype strains. CONCLUSIONS: The proportion of the rotavirus genotypes observed was not different from that in other areas of Brazil. This study is the first genotyping of rotavirus in the Western Brazilian Amazon.

scholarly journals Isolation, immunochemical demonstration of field strains of porcine group A rotaviruses and electrophoretic analysis of RNA segments of group A and C rotaviruses

2012 ◽  
Vol 51 (No. 5) ◽  
pp. 288-295 ◽  
Author(s):  
R. Smitalova ◽  
L. Rodak ◽  
I. Psikal ◽  
B. Smid
Keyword(s):  
Cell Line ◽  
Electrophoretic Analysis ◽  
Elisa Test ◽  
Farm Animals ◽  
Economic Losses ◽  
Rt Pcr ◽  
Group C Rotavirus ◽  
Group A Rotaviruses ◽  
Group A ◽  
Virus Isolates

Rotaviruses are major cause of acute diarrhea in animals and humans which can result in huge economic losses in farm animals including pigs. We collected 195 samples of feces of diarrhoeic animals. Rotavirus was demonstrated by electron microscopy using the method of negative staining in 27 samples and by ELISA test using monoclonal antibodies to the group antigen VP6 in 44 samples. Nine samples were selected for virus isolation. Three virus isolates (P375/4, P410/4 and P646/1) were successfully adapted to growth in cell line MA-104. These isolates were allocated to group A rotaviruses based on ELISA, immunoperoxidase test and electropherotype analysis. Electropherotype analysis demonstrated changes during passage in cell line in two of the three isolates. The selected sample P543/1 proved negative in ELISA in a fecal sample. Electropherotype analysis of this sample revealed a “longer” electropherotype profile. The profile was suggestive of group C rotavirus. Rotavirus group C was confirmed by RT-PCR and by sequence analysis in this sample.

Detection of mammalian-like group a rotavirus in diarrhoeic poultry using RNA-PAGE In Kerala, India

2018 ◽  
Author(s):  
Rinsha Balan ◽  
M. Mini ◽  
P. M. Priya ◽  
Siju Joseph ◽  
Surya Sankar
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Virus Detection ◽  
Migration Pattern ◽  
Polyacrylamide Gel ◽  
Rota Virus ◽  
Poultry Birds ◽  
Faecal Samples ◽  
Group A ◽  
Different Parts

The present work was conducted to determine the prevalence of rotavirus infection among poultry birds having diarrhoea. A total of 143 faecal samples were collected from different parts of Kerala and screened for the presence of rotavirus using RNA polyacrylamide gel electrophoresis (RNA-PAGE). Out of 143 samples, 5 (3.49%) were found to be positive in RNA PAGE with a migration pattern 4:2:3:2 of a mammalian-like electropherogroup A rotavirus. The study records the first evidence of rota virus detection from avian species in Kerala.

scholarly journals Incidence of group A rotavirus in urban and rural areas of the city of Londrina-Brazil, from 1995 to 1997

2001 ◽  
Vol 44 (3) ◽  
pp. 257-261 ◽  
Author(s):  
Ângelo Cesar Meneghetti ◽  
Andrea Maria Bolognini ◽  
Flávio Lauretti ◽  
Rosa Elisa Carvalho Linhares ◽  
Norma Santos ◽  
...  
Keyword(s):  
Rural Areas ◽  
Gel Electrophoresis ◽  
Diarrheal Disease ◽  
Polyacrylamide Gel Electrophoresis ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Rt Pcr ◽  
Group A ◽  
Urban And Rural Areas ◽  
The City

Rotaviruses are common pathogens and the causal agents of acute diarrhea among children and young animals. The involvement of rotavirus in human diarrheal disease among population of urban and rural areas of the city of Londrina, Parana was evaluated. Nine hundred and five fecal specimens from persons with diarrhea were studied, being 686 and 219 from urban and rural areas, respectively. Thirty-eight samples (4,2%) were positive for rotavirus by polyacrylamide gel electrophoresis of viral RNA and latex agglutination test of which 36 were from urban and two from rural areas. Out of the positive specimens, 17 strains were further characterized by RT-PCR typing assay, resulting in 16 strains of G1 genotype while one sample was found to be a mixture of G1 and G3 genotypes.

scholarly journals Comparison of electron microscopy, enzyme-linked immunosorbent assay and latex agglutination for the detection of bovine rotavirus in faeces

1997 ◽  
Vol 68 (3) ◽  
Author(s):  
M. De Beer ◽  
I. Peenze ◽  
V.M. Da Costa Mendes ◽  
A.D. Steele
Keyword(s):  
Electron Microscopy ◽  
Gold Standard ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Latex Agglutination ◽  
The Other ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Rotavirus ◽  
Group A Rotaviruses ◽  
Group A

The performance characteristics of 2 enzyme immunoassays (ELISAs) and 4 latex agglutination assays (LXs) were evaluated for the detection of bovine rotavirus in faecal specimens of young calves with diarrhoea. A total of 26 specimens from calves less than 5 months of age were examined with different commercial assays and compared with electron microscopy (EM) as the gold standard and with polyacrylamide gel electrophoresis (PAGE) for the detection of atypical, non-group A rotaviruses. In the 2nd study, EIA (Dako) and LX (Murex), the assays of choice, were used to analyse 97 further faecal specimens from calves with diarrhoea. The ELISAs proved to be the most sensitive compared with the other tests used. The EM and PAGE are 100 % specific although slightly less sensitive than the commercial assays. The results show that all the commercial assays can accurately detect rotavirus in the stools of calves with gastroenteritis, although the suitability and choice of assay will depend upon the requirements of individual laboratories.

scholarly journals Sequence analysis of the guanylyltransferase (VP3) of group A rotaviruses

2004 ◽  
Vol 85 (4) ◽  
pp. 929-932 ◽  
Author(s):  
Jonathan P. Cook ◽  
Malcolm A. McCrae
Keyword(s):  
Mammalian Species ◽  
Sequence Divergence ◽  
Amino Acid Sequences ◽  
Rt Pcr ◽  
Human Virus ◽  
Sequence Identity ◽  
Group A Rotaviruses ◽  
Group A ◽  
Mammalian Origin ◽  
Virus Isolates

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 identity. The exceptions were avian virus isolates, which diverged ∼45 % from those of mammalian origin, and the human virus isolates DS1 and 69M, which showed much closer (over 90 %) identity to viruses of bovine origin, suggesting that these human isolates may have undergone recent reassortment events with a bovine virus. Analysis of the sequences for a putative enzymic active site has revealed that the KXTAMDXEXP and KXXGNNH motifs around amino acids 385 and 545, respectively, are conserved across both group A and C rotaviruses.

Rotavirus electropherotypes in Malaysian children

1992 ◽  
Vol 38 (9) ◽  
pp. 996-999 ◽  
Author(s):  
K. L. Yap ◽  
Y. H. Wong ◽  
C. M. Khor ◽  
Y. E. Ooi
Keyword(s):  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Older Children ◽  
Migration Patterns ◽  
Faecal Samples ◽  
Group A Rotaviruses ◽  
Group A ◽  
Short Pattern ◽  
Predominant Strain

A 12-month study was carried out on the molecular epidemiology of rotavirus in urban and suburban Malaysian children. Analysis of faecal samples from 973 hospitalized diarrhoeic children by polyacrylamide gel electrophoresis detected 268 rotaviruses (28%). All isolates were group A rotaviruses, which produced 22 electropherotypes: 16 (91.5%) with long RNA migration patterns and 6 (8.5%) with short patterns. One of the long-pattern electropherotypes was the predominant strain (71.1% of the total electropherotypes) isolated during this study. Although 3 other strains were detected sporadically over the study period, 16 others were present only during the first 7 months and 2 others were confined to the last 5 months. Long- and short-pattern electropherotypes were found to co-circulate extensively. There was a significant association of short-pattern electropherotypes with infection in older children. In addition, the prevalence of vomiting and mean duration of diarrhoea were significantly associated with different electropherotypes. Key words: rotavirus electropherotypes.

scholarly journals Dual Infection of Gnotobiotic Calves with Bovine Strains of Group A and Porcine-Like Group C Rotaviruses Influences Pathogenesis of the Group C Rotavirus

1999 ◽  
Vol 73 (11) ◽  
pp. 9284-9293 ◽  
Author(s):  
K. O. Chang ◽  
P. R. Nielsen ◽  
L. A. Ward ◽  
L. J. Saif
Keyword(s):  
Villous Atrophy ◽  
Polyacrylamide Gel Electrophoresis ◽  
Dual Infection ◽  
The United States ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Group C Rotavirus ◽  
Bovine Strain ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT There is serological evidence that bovine group C rotaviruses exist in the United States, but there are no reports of their isolation. Ninety fecal samples from calves with diarrhea, 81 samples from adult cows with diarrhea (winter dysentery), and 20 fecal samples from healthy adult cows were tested for group C rotaviruses by polyacrylamide gel electrophoresis, immune electron microscopy, and reverse transcription-PCR (RT-PCR). Three samples from adult cow diarrhea cases were positive only by RT-PCR, and a group C rotavirus was isolated from a positive sample in monkey kidney (MA104) cells (WD534tc/C). Genetically and serologically, the WD534tc/C strain was more closely related to the Cowden porcine group C strain than to the Shintoku bovine strain. Because the original cow feces also contained a group A rotavirus (detected after passage in cell culture), we hypothesized that such dual-rotavirus infections might play a role in the pathogenesis and host adaptation of rotaviruses. Thus, we examined the pathogenesis of WD534tc/C alone or combined with virulent (IND/A) or attenuated (NCDV/A) bovine group A rotaviruses in gnotobiotic calves. WD534tc/C alone induced diarrhea without (or with limited) virus shedding in inoculated calves (n = 3). In contrast, all calves coinfected with WD534tc/C and IND/A (n = 2) developed diarrhea and shed both viruses, whereas calves coinfected with WD534tc/C and NCDV/A (n= 3) developed diarrhea but did not shed either virus. Infection with WD534tc/C or NCDV/A alone caused only mild villous atrophy (jejunum and/or ileum), whereas dual infection with both viruses induced lesions throughout the small intestine. Although IND/A alone caused villous atrophy, more-widespread small intestinal lesions occurred in calves coinfected with WD534tc/C and IND/A. In conclusion, coinfection of calves with group A rotaviruses enhanced fecal shedding of a bovine group C rotavirus and the extent of histopathological lesions in the small intestines. Thus, our findings suggest a potential novel hypothesis involving dual infections for the adaptation of heterologous rotaviruses to new host species.

scholarly journals Suspected zoonotic transmission of rotavirus group A in Danish adults

2011 ◽  
Vol 140 (6) ◽  
pp. 1013-1017 ◽  
Author(s):  
S. E. MIDGLEY ◽  
C. K. HJULSAGER ◽  
L. E. LARSEN ◽  
G. FALKENHORST ◽  
B. BÖTTIGER
Keyword(s):  
Genetic Relatedness ◽  
Phylogenetic Analyses ◽  
Geographical Area ◽  
Nucleotide Sequences ◽  
Zoonotic Transmission ◽  
Adult Patients ◽  
Epidemiological Features ◽  
Group A Rotaviruses ◽  
Group A ◽  
Stool Samples

SUMMARYGroup A rotaviruses infect humans and a variety of animals. In July 2006 a rare rotavirus strain with G8P[14] specificity was identified in the stool samples of two adult patients with diarrheoa, who lived in the same geographical area in Denmark. Nucleotide sequences of the VP7, VP4, VP6, and NSP4 genes of the identified strains were identical. Phylogenetic analyses showed that both Danish G8P[14] strains clustered with rotaviruses of animal, mainly, bovine and caprine, origin. The high genetic relatedness to animal rotaviruses and the atypical epidemiological features suggest that these human G8P[14] strains were acquired through direct zoonotic transmission events.

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