Comparison of different RT-qPCR assays for the detection of human and bovine group A rotaviruses and characterization by sequences analysis of genes encoding VP4 and VP7 capsid proteins

Group A rotaviruses belong to the Reoviridae virus family and are classified into G and P genotypes based on the outer capsid proteins VP7 and VP4, respectively [...]

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

Journal of Virology ◽

10.1128/jvi.76.13.6596-6601.2002 ◽

2002 ◽

Vol 76 (13) ◽

pp. 6596-6601 ◽

Cited By ~ 174

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.

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DNA Damage-Induced Neurodegeneration in Accelerated Ageing and Alzheimer’s Disease

International Journal of Molecular Sciences ◽

10.3390/ijms22136748 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6748

Author(s):

Heling Wang ◽

Sofie Lautrup ◽

Domenica Caponio ◽

Jianying Zhang ◽

Evandro F. Fang

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Dna Repair ◽

Werner Syndrome ◽

Accelerated Ageing ◽

Genes Encoding ◽

Group A ◽

Group B ◽

Novel Therapeutic Strategies ◽

Energy Deprivation

DNA repair ensures genomic stability to achieve healthy ageing, including cognitive maintenance. Mutations on genes encoding key DNA repair proteins can lead to diseases with accelerated ageing phenotypes. Some of these diseases are xeroderma pigmentosum group A (XPA, caused by mutation of XPA), Cockayne syndrome group A and group B (CSA, CSB, and are caused by mutations of CSA and CSB, respectively), ataxia-telangiectasia (A-T, caused by mutation of ATM), and Werner syndrome (WS, with most cases caused by mutations in WRN). Except for WS, a common trait of the aforementioned progerias is neurodegeneration. Evidence from studies using animal models and patient tissues suggests that the associated DNA repair deficiencies lead to depletion of cellular nicotinamide adenine dinucleotide (NAD+), resulting in impaired mitophagy, accumulation of damaged mitochondria, metabolic derailment, energy deprivation, and finally leading to neuronal dysfunction and loss. Intriguingly, these features are also observed in Alzheimer’s disease (AD), the most common type of dementia affecting more than 50 million individuals worldwide. Further studies on the mechanisms of the DNA repair deficient premature ageing diseases will help to unveil the mystery of ageing and may provide novel therapeutic strategies for AD.

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A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879400600207 ◽

1994 ◽

Vol 6 (2) ◽

pp. 175-181 ◽

Cited By ~ 16

Author(s):

A. Lucchelli ◽

S. Y. Kang ◽

M. K. Jayasekera ◽

A. V. Parwani ◽

D. H. Zeman ◽

...

Keyword(s):

Monoclonal Antibodies ◽

South Dakota ◽

Dairy Herd ◽

Dairy Calves ◽

Enzyme Linked Immunosorbent Assay ◽

Positive Stool ◽

Field Samples ◽

Group A Rotaviruses ◽

Beef Herds ◽

Group A

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.

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Regulation of Polysaccharide Utilization Contributes to the Persistence of Group A Streptococcus in the Oropharynx

Infection and Immunity ◽

10.1128/iai.00081-07 ◽

2007 ◽

Vol 75 (6) ◽

pp. 2981-2990 ◽

Cited By ~ 16

Author(s):

Samuel A. Shelburne ◽

Nnaja Okorafor ◽

Izabela Sitkiewicz ◽

Paul Sumby ◽

David Keith ◽

...

Keyword(s):

Parental Strain ◽

Group A Streptococcus ◽

Transcriptional Regulator ◽

Human Saliva ◽

Transcript Levels ◽

Expression Of Genes ◽

Rich Media ◽

Genes Encoding ◽

Group A ◽

Mutant Derivative

ABSTRACT Group A Streptococcus (GAS) genes that encode proteins putatively involved in polysaccharide utilization show growth phase-dependent expression in human saliva. We sought to determine whether the putative polysaccharide transcriptional regulator MalR influences the expression of such genes and whether MalR helps GAS infect the oropharynx. Analysis of 32 strains of 17 distinct M protein serotypes revealed that MalR is highly conserved across GAS strains. malR transcripts were detectable in patients with GAS pharyngitis, and the levels increased significantly during growth in human saliva compared to the levels during growth in glucose-containing or nutrient-rich media. To determine if MalR influenced the expression of polysaccharide utilization genes, we compared the transcript levels of eight genes encoding putative polysaccharide utilization proteins in the parental serotype M1 strain MGAS5005 and its ΔmalR isogenic mutant derivative. The transcript levels of all eight genes were significantly increased in the ΔmalR strain compared to the parental strain, especially during growth in human saliva. Following experimental infection, the ΔmalR strain persistently colonized the oropharynx in significantly fewer mice than the parental strain colonized, and the numbers of ΔmalR strain CFU recovered were significantly lower than the numbers of the parental strain CFU recovered. These data led us to conclude that MalR influences the expression of genes putatively involved in polysaccharide utilization and that MalR contributes to the persistence of GAS in the oropharynx.

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

ISRN Virology ◽

10.5402/2013/179871 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 2

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.

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Survey of Human Group C Rotaviruses in Japan during the Winter of 1992 to 1993

Journal of Clinical Microbiology ◽

10.1128/jcm.36.1.6-10.1998 ◽

1998 ◽

Vol 36 (1) ◽

pp. 6-10 ◽

Cited By ~ 27

Author(s):

Mitsutaka Kuzuya ◽

Ritsushi Fujii ◽

Masako Hamano ◽

Masao Yamada ◽

Kuniko Shinozaki ◽

...

Keyword(s):

Large Scale ◽

Blot Hybridization ◽

Human Group ◽

Dot Blot ◽

Passive Hemagglutination ◽

Group A Rotaviruses ◽

Group A ◽

Outer Capsid ◽

Very High ◽

Vp7 Gene

Fecal specimens from patients with acute diarrhea were collected from 10 prefectures in Japan over a 6-month period (November 1992 to April 1993), and the specimens that were negative for human group A rotaviruses were screened for the presence of human group C rotaviruses (CHRVs) by the reverse passive hemagglutination test. Of 784 specimens examined, 53 samples (6.8%) that were collected in 7 of 10 prefectures were positive for CHRV, indicating that CHRVs are widely distributed across Japan. Most of the CHRV isolates were detected in March and April, and CHRVs mainly prevailed in children ages 3 to 8 years. The genome electropherotypes of eight strains isolated in five individual prefectures were surprisingly similar to each other and were different from those of CHRV strains isolated to date. The outer capsid glycoprotein (VP7) gene homologies of the isolates retrieved in 1993 were subsequently analyzed by the dot blot hybridization method. As a result, the VP7 genes of the isolates revealed very high levels of homology not only with each other but also with the VP7 gene of the OK118 strain isolated in 1988. These results suggest that a large-scale outbreak of CHRV occurred during the winter of 1992 and 1993 in Japan.

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Suspected zoonotic transmission of rotavirus group A in Danish adults

Epidemiology and Infection ◽

10.1017/s0950268811001981 ◽

2011 ◽

Vol 140 (6) ◽

pp. 1013-1017 ◽

Cited By ~ 21

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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