Development of ELISA kits for antibodies against avian reovirus using the σC and σB proteins expressed in the methyltropic yeast Pichia pastoris

Yang, Zijiang; Wang, Chi-Young; Lee, Long-Huw; Chuang, Kui-Hao; Lien, Yi-Yang; Yin, Hao; Tong, Dewen; Xu, Xiaoxiao; Liu, Hung-Jen

doi:10.1016/j.jviromet.2009.07.009

Cited by 17 publications

(9 citation statements)

References 39 publications

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“…It has also been suggested that it represents the target for type-specific neutralizing antibodies. The σB protein, encoded by the S3 gene [10,19], carries group-specific neutralizing epitopes [20]. Recently, the ARV σA protein, encoded by the S2 gene [16], has been identified as a double-stranded RNA binding protein that may be involved in interferon resistance [21].…”

Section: Introductionmentioning

confidence: 99%

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: Introductionmentioning

confidence: 99%

Molecular characterization of avian reovirus isolates in Tunisia

Kort

Bourogâa

Gribaa

et al. 2013

Virol J

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“…To establish an ELISA for detection of the PCV2 Cap(d41) antibody, the optimal dilutions of antigen and serum were determined by checker board titration with PCV2-positive and -negative swine sera [24]. For expression of Cap(d41) in E. coli, the PCV2 Cap gene with a deletion of the first 123 bp was amplified by PCR using prime pair PCV2-Cap(d41)-XhoI and PCV2-Cap(d41)-PstI (Table 1) and subcloned into the expression vector pET32a.…”

Section: Comparison Of the Expression Levels Of The Pcv2 Cap(d41) Promentioning

confidence: 99%

Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine

Chen

Chang

et al. 2020

Vet Res

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To increase expression levels of the PCV2 Cap(d41) protein, novel baculovirus surface display vectors with multiple expression cassettes were constructed to create recombinant baculoviruses BacSC-Cap(d41), BacDD-2Cap(d41), BacDD-3Cap(d41), and BacDD-4Cap(d41). Our results reveal that the recombinant baculovirus BacDD-4Cap(d41) was able to express the highest levels of Cap(d41) protein. Optimum conditions for expressing the PCV2 Cap(d41) protein were determined, and our results show that 107 of Sf-9 infected with the recombinant baculovirus BacDD-4Cap(d41) at an MOI of 5 for 3 days showed the highest level of protein expression. Mice immunized with the 4Cap(d41) vaccine which was prepared from the recombinant baculovirus-infected cells (107) elicited higher ELISA titers compared to the Cap (d41) vaccine. The 4Cap(d41) vaccine could elicit anti-PCV2 neutralizing antibodies and IFN-γ in mice, as confirmed by virus neutralization test and IFN-γ ELISA. Moreover, the swine lymphocyte proliferative responses indicated that the 4Cap(d41) vaccine was able to induce a clear cellular immune response. Flow cytometry analysis showed that the percentage of CD4+ T cells and CD4+/CD8+ ratio was increased significantly in SPF pigs immunized with the 4Cap(d41) vaccine. Importantly, the 4Cap(d41) vaccine induced an IFN-γ response, further confirming that its effect is through cellular immunity in SPF pigs. An in vivo challenge study revealed that the 4Cap(d41) and the commercial vaccine groups significantly reduce the viral load of vaccinated pigs as compared with the CE negative control group. Taken together, we have successfully developed a 4Cap(d41) vaccine that may be a potential subunit vaccine for preventing the disease associated with PCV2 infections.

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“…Algumas classificações de diferentes isolados do vírus ARV foram estabelecidas por sorologia que utilizaram testes de neutralização, agrupando, então, as estirpes em sorotipos (HIERONYMUS; VILLEGAS; KLEVEN, 1983;WOOD et al, 1980). As proteínas de ARV que induzem a produção de anticorpos neutralizantes e que, por conseguinte, estão envolvidas com inibição de infectividade viral, são σC, σB e λC (YANG et al, 2010). Porém, a considerável heterogeneidade dessas proteínas tem dificultado a classificação das estirpes de ARV por meio da caracterização sorológica, resultando em grande número de reações cruzadas em testes de neutralização (BENAVENTE; MARTÍNEZ-COSTAS, 2007;CLARK et al, 1990).…”

Section: Classificação Dos Isolados Viraisunclassified

Reovirose aviária: um panorama

Assunção

Palka

Pavoni

2018

Rev. Educ. Cont. Med. Vet. Zootec.

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A avicultura é um setor de grande importância na economia brasileira tanto pelo aumento do consumo interno quanto pelo crescimento na exportação de carne de frango. A modernização da atividade avícola permitiu a criação adensada de animais, facilitando, no entanto, a rápida disseminação de patógenos que reduzem os índices de produtividade dos plantéis. Nesta revisão, é destacado o reovírus aviário, importante agente de artrite em aves que apresenta distribuição mundial. As aves acometidas apresentam redução no ganho de peso devido à dificuldade de locomoção. Além da artrite, o vírus pode estar relacionado a uma variedade de condições patológicas, como distúrbios entéricos e respiratórios, hepatite e miocardite. A principal forma de prevenção e controle é a vacinação do plantel. No entanto, devido à grande variabilidade genética do reovírus aviário, a vacina utilizada pode não ser eficiente contra estirpes que circulam no campo. O artigo traz uma visão geral sobre a biologia do vírus, sua variabilidade e propostas de classificação dos isolados, patologia da doença e diagnóstico da infecção.

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Development of ELISA kits for antibodies against avian reovirus using the σC and σB proteins expressed in the methyltropic yeast Pichia pastoris

Cited by 17 publications

References 39 publications

Molecular characterization of avian reovirus isolates in Tunisia

Molecular characterization of avian reovirus isolates in Tunisia

Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine

Reovirose aviária: um panorama

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