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 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 Characterization of rotavirus electropherotypes excreted by symptomatic and asymptomatic infants

1991 ◽  
Vol 106 (1) ◽  
pp. 189-198 ◽  
Author(s):  
J. Fernández ◽  
A. M. Sandino ◽  
J. Pizarro ◽  
L. F. Avendaño ◽  
J. M. Pizarro ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Mixed Infections ◽  
Migration Patterns ◽  
Virulent Strains ◽  
Human Rotavirus ◽  
Stool Samples

SUMMARYHuman rotavirus isolates from 1100 stool samples were analyzed by polyacrylamide gel electrophoresis, and 48 different migration patterns were detected. Heterogeneity in the migration of segment 10 was observed in both long and short electropherotypes in which three long and two short patterns were identified. In spite of these variations all short and long electropherotypes were subgrouped by enzyme immunoassay as subgroups I and II respectively. Mixed infections were detected in 17% of cases and the subgrouping correlated with the corresponding electropherotypes. The same electropherotypes were present in severe, mild and asymptomatic cases and no electropherotype was particularly associated with greater virulence. Furthermore, the electropherotypes isolated from nosocomial asymptomatic cases were the same as those detected from those admitted with severe diarrheoa. It seems unlikely that electropherotyping can be used to identify more virulent strains of rotavirus.

scholarly journals Molecular characterization of group A bovine rotavirus in southeastern and central-western Brazil, 2009-2010

2012 ◽  
Vol 32 (3) ◽  
pp. 237-242 ◽  
Author(s):  
Fernanda D.F. Silva ◽  
Fábio Gregori ◽  
Ana C.S. Gonçalves ◽  
Samir Issa Samara ◽  
Maria G. Buzinaro
Keyword(s):  
Molecular Characterization ◽  
Polyacrylamide Gel Electrophoresis ◽  
Genotypic Diversity ◽  
The State ◽  
Mato Grosso ◽  
Bovine Rotavirus ◽  
Prophylactic Measures ◽  
Beef Herds ◽  
Group A

Rotavirus is an important cause of neonatal diarrhea in humans and several animal species, including calves. A study was conducted to examine 792 fecal samples collected from calves among 65 dairy and beef herds distributed in two of Brazil's major livestock producing regions, aiming to detect the occurrence of rotavirus and perform a molecular characterization of the rotavirus according to G and P genotypes in these regions. A total of 40 (5.05%) samples tested positive for rotavirus by the polyacrylamide gel electrophoresis (PAGE) technique. The molecular characterization was performed by multiplex semi-nested RT-PCR reactions, which indicated that the associations of genotypes circulating in herds in Brazil's southeastern region were G6P[11], G10P[11], G[-]P[5] + [11], G[-]P[6] in the state of São Paulo and G6P[11], G8P[5], G11P[11], G10P[11] in the state of Minas Gerais. In the central-western region, the genotypes G6P[5] + [11], G6P[5], G8P[-], G6P[11], G [-] P[1], G[-] P[11], and G[-] P[5] were detected in the state of Goiás, while the genotypes G6P[5], G8[P11], G6[P11], G8[P1], G8[P5], G6[P1] were circulating in herds in the state of Mato Grosso do Sul. The genotypic diversity of bovine rotavirus found in each region under study underlines the importance of characterizing the circulating samples in order to devise the most effective prophylactic measures.

scholarly journals Molecular Characterization of Rotavirus in Ireland: Detection of Novel Strains Circulating in the Population

2000 ◽  
Vol 38 (9) ◽  
pp. 3370-3374 ◽  
Author(s):  
F. O'Halloran ◽  
M. Lynch ◽  
B. Cryan ◽  
H. O'Shea ◽  
S. Fanning
Keyword(s):  
Nucleic Acid ◽  
Molecular Characterization ◽  
Molecular Diagnostics ◽  
Polyacrylamide Gel ◽  
Mixed Infections ◽  
Positive Stool ◽  
Stool Samples ◽  
Institute Of Technology

A collection of three hundred thirty rotavirus-positive stool samples from children with diarrhea in the southern and eastern regions of Ireland between 1997 and 1999 were submitted to the Molecular Diagnostics Unit of the Cork Institute of Technology, Cork, Ireland, for investigation. These strains were characterized by several methods, including polyacrylamide gel electropherotyping and G and P genotyping. A subset of the G types was confirmed by nucleic acid sequencing. The most prevalent types found in this collection included G1P[8] (n = 106; 32.1%), G2P[4] (n = 94; 28.5%), and G4P[8] (n = 37; 11.2%). Novel strains were also detected, including G1P[4] (n = 19; 5.8%), and G4P[4] (n = 2; 0.6%). Interestingly, mixed infections accounted for 18.8% (n = 62) of the total collection, with only 3% (n = 10) which were not G and/or P typeable. Significantly, six G8 and five G9 strains were identified as part of mixed infections. These strains have not previously been identified in Irish children, suggesting a greater diversity in rotavirus strains currently circulating in Ireland.

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 Characterization of rotavirus P genotypes circulating among paediatric inpatients in Northern Brazil

1999 ◽  
Vol 41 (3) ◽  
pp. 165-170 ◽  
Author(s):  
Joana D’Arc P. MASCARENHAS ◽  
Rosa Helena P. GUSMÃO ◽  
Célia R. M. BARARDI ◽  
Fernanda L. PAIVA ◽  
Cláudia O. SIMÕES ◽  
...  
Keyword(s):  
Type Strain ◽  
The Other ◽  
Rt Pcr ◽  
Other Hand ◽  
Positive Stool ◽  
Northern Brazil ◽  
Stool Samples ◽  
Gene 4 ◽  
Type Infection

Between November 1992 and August 1993, twenty-eight rotavirus-positive stool samples obtained from paediatric inpatients in Belém, Brazil, aged less than four years, were tested by RT-PCR to determine the P genotype specificities. With the exception of 7 non-diarrhoeic children, all patients were either diarrhoeic at admission or developed diarrhoea while in hospital. Rotavirus strains with the gene 4 alleles corresponding to P1B[4] and P1A[8] types (both of which bearing G2 specificity) predominated, accounting for 78.6% of the strains. While only one P2A[6] type strain - with (mixed) G1 and 4 type specificities - was detected, the gene 4 allele could not be identified in 4 (14.3%) of the strains. Most (81%) of the specimens were obtained from children during their first 18 months of life. Rotavirus strains bearing single P1B[4] type-specificity were identified in both diarrhoeic (either nosocomial, 28.6% or community-acquired diarrhoea, 28.6%) and non-diarrhoeic (42.8%) children. P1A[8] gene 4 allele, on the other hand, was detected only among diarrhoeic children, at rates of 57.1% and 42.9% for nosocomial- and- community acquired diarrhoea, respectively. Mixed P1A[8],1B[4] type infection was identified in only one case of community-acquired diarrhoea.

scholarly journals Molecular Characterization of VP6 Genes of Human Rotavirus Isolates: Correlation of Genogroups with Subgroups and Evidence of Independent Segregation

2002 ◽  
Vol 76 (13) ◽  
pp. 6596-6601 ◽  
Author(s):  
Miren Iturriza Gómara ◽  
Cecilia Wong ◽  
Sandra Blome ◽  
Ulrich Desselberger ◽  
Jim Gray
Keyword(s):  
Molecular Characterization ◽  
Rt Pcr ◽  
Genetic Lineages ◽  
Reverse Transcription Pcr ◽  
Human Rotavirus ◽  
Independent Segregation ◽  
Genes Encoding ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT 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 genogroups. Two genetic lineages were distinguished within genogroup I, consisting of strains serologically characterized as SG I, and three genetic lineages were distinguished within genogroup II, composed of strains serologically characterized as SG II, SG I + II, and SG non-I, non-II. Subgrouping of rotaviruses by means of serological methods may result in strains not being assigned the correct SG or in a failure of strains to subgroup. Molecular characterization of the SG-defining region of VP6 provided evidence for independent segregation of the rotavirus genes encoding VP4, VP6, and VP7.

Prothrombin Metz : Purification and Characterization of a Variant of Human Prothrombin

1979 ◽  
Author(s):  
M Ribieto ◽  
J Elion ◽  
D Labie ◽  
F Josso
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Xa ◽  
Polyacrylamide Gel ◽  
Cleavage Pattern ◽  
Purification And Characterization ◽  
Double Immunodiffusion ◽  
The Family

For the purification of the abnormal prothrombin (Pt Metz), advantage has been taken of the existence in the family of three siblings who, being double heterozygotes for Pt Metz and a hypoprothrombinemia, have no normal Pt. Purification procedures included barium citrate adsorption and chromatography on DEAE Sephadex as for normal Pt. As opposed to some other variants (Pt Barcelona and Madrid), Pt Metz elutes as a single symetrical peak. By SDS polyacrylamide gel electrophoresis, this material is homogeneous and appears to have the same molecular weight as normal Pt. Comigration of normal and abnormal Pt in the absence of SDS, shows a double band suggesting an abnormal charge for the variant. Pt Metz exhibits an identity reaction with the control by double immunodiffusion. Upon activation by factor Xa, Pt Metz can generate amydolytic activity on Bz-Phe-Val-Arg-pNa (S2160), but only a very low clotting activity. Clear abnormalities are observed in the cleavage pattern of Pt Metz when monitored by SDS gel electrophoresis. The main feature are the accumulation of prethrombin l (Pl) and the appearance of abnormal intermediates migrating faster than Pl.

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