Detection of avian reovirus RNA and comparison of a portion of genome segment S3 by polymerase chain reaction and restriction enzyme fragment length polymorphism

Lee, Long Huw; Shien, Jui Hung; Shieh, Happy K.

doi:10.1016/s0034-5288(98)90020-0

Cited by 27 publications

(6 citation statements)

References 15 publications

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“…Serological classification of ARV strains has not been successful because of high cross-reactivity of the neutralizing antibodies [33]. Nowadays, genotypic classification of ARV strains is performed using RT-PCR in combination with phylogenic analysis or other molecular techniques, such as RFLP [7,26]. All phylogenic studies have classified ARV isolates into various groups and lineages; however, meta-analytically, there were no identical patterns.…”

Section: Discussionmentioning

confidence: 99%

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Molecular characterization of avian reovirus isolates in Tunisia

Kort

Bourogâa

Gribaa

et al. 2013

Virol J

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BackgroundGenotype analyses of avian reoviruses isolated from organ samples collected from chickens with suspicious clinical symptoms, between 1997–2008, was based on sequences for both σC and σB genes and aligned with those published in the Genbank, making it possible to carry out studies of molecular classification and relationships.MethodsThe full length of the known variable protein σC and part of the σB encoding genes, were amplified with RT-PCR, using conserved primers. PCR products were sequenced and the sequences were analyzed and aligned with avian reovirus sequences from the Genbank database.ResultsThe sequences of σC-encoding genes of all the isolated strains indicated their close relationship with the American, Chinese and Indian strains. Taking the American strain S1133 as a reference, the two Tunisian isolates 97.1 and 97.2 showed some nucleotide substitutions. For isolate 97.1, the substitution was silent whereas for strain 97.2 the mutation was at the first position of the corresponding codon and induced the substitution of the amino acid encoded. For the σB-encoding gene, the sequences of the Tunisian strains showed mutations at positions two or three of the corresponding codons, inducing substitutions of amino acids at these positions. The phylogenic trees based on σC and σB encoding genes indicated closer relationship between Tunisian, American and Taiwanese isolates of genotype I.ConclusionOur study describes the genotype of avian reoviruses that are not yet well characterized genetically. The characterization and classification of these viruses might be significant for understanding the epidemiology of malabsorption syndrome and viral arthritis, and improving our knowledge of the genotype of strains circulating in Tunisian flocks. Furthermore, the study of their variable pathogenicity could be extremely important in the choice of the appropriate vaccine strain to control disease.

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Section: Discussionmentioning

confidence: 99%

“…The vaccine strain (Nobilis S1133) was propagated in CEF cells and treated as described above. Viral RNA was extracted from supernatants using Trizol (Gibco Brl) as per the procedural modification described by Lee et al [26]. Briefly, 1ml of Trizol was incubated with 300 μl of clarified supernatant for 5min, at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of avian reovirus isolates in Tunisia

Kort

Bourogâa

Gribaa

et al. 2013

Virol J

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“…Several methods such as virus isolation, dot-blot hybridization, reverse transcriptase polymerase chain reaction (RT-PCR), RT-PCR restriction fragment length polymorphism, real-time RT-PCR, and enzyme-linked immunosorbent assay have been used in the diagnosis of ARV infection (Liu & Giambrone, 1997;Xie et al, 1997Xie et al, , 2010Lee et al, 1998;Yin & Lee, 1998;Liu et al, 1999;Guo et al, 2011). However, virus isolation is timeconsuming and the other methods require specialized equipment, which makes them difficult to apply under field conditions for rapid detection of ARV.…”

Section: Introductionmentioning

confidence: 99%

Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of avian reovirus

et al. 2012

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“…Additionally, virus isolation [ 11 ], immunofluorescent staining [ 12 ] and immunoperoxidase histochemistry [ 13 , 14 ] offer the straight detection of viral antigens in tendon tissues. The methods for detection of ARV RNA from the cell culture [ 15 ] or from both cell cultures and tendon specimens [ 16 ] have been developed to provide a sensitive tool for the laboratory diagnosis. The serological diagnosis methods such as ELISA were also used for detection of serum antibodies in large number of samples simultaneously [ 17 – 20 ].…”

Section: Introductionmentioning

confidence: 99%

In vitro study on role of σB protein in avian reovirus pathogenesis

et al. 2018

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Avian reoviruses, members of Orthoreovirus genus was known to cause diseases like tenosynovitis, runting-stunting syndrome in chickens. Among eight structural proteins, the proteins of S-class are mainly associated with viral arthritis but the significance of σB protein in arthritis is not established till date. In this infection pathological condition together with infection of joints often leads to arthritis because joints consists of cartilage which forms lubricating surface between two bones, and has limited metabolic, replicative and repair capacity.To establish the role of σB protein in arthritis, an in-vitro microarray study was conducted consisting four groups viz. virus infected and control; pDsRed-Express-N1-σB and empty pDs-Red transfected, CEF cells. With cut-off value as FC ≥2, p value <0.05, 6709 and 4026 numbers of DEGs in virus and σB, respectively were identified. The Ingenuity Pathway Analysis gave an idea about the involvement of σB protein in “osteoarthritis pathway”, which was activated with z-score with 3.151. The pathway “Role of IL-17A in arthritis pathway” was also enriched with –log (p-value) 1.64. Among total 122 genes involved in osteoarthritis pathway, 28 upregulated and 11 downregulated DEGs were common to both virus and σB treated cells. Moreover, 14 upregulated and 7 downregulated were unique in σB transfected cells. Using qRT-PCR for IL-1B, BMP2, SMAD1, SPP1 genes, the microarray data was validated. We concluded that during ARV infection σB protein, if not fully partially leads to molecular alteration of various genes of host orchestrating the different molecular pattern in joints, leading to tenosynovitis syndrome.

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Detection of avian reovirus RNA and comparison of a portion of genome segment S3 by polymerase chain reaction and restriction enzyme fragment length polymorphism

Cited by 27 publications

References 15 publications

Molecular characterization of avian reovirus isolates in Tunisia

Molecular characterization of avian reovirus isolates in Tunisia

Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of avian reovirus

In vitro study on role of σB protein in avian reovirus pathogenesis

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