Expression and immunogenicity analysis of the capsid proteins of porcine circovirus types 2 to 4
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Antiserum to electrophoretically-separated capsid protein of grapevine virus B (GVB) was produced. After easy and effective elimination of antibodies cross-reactive with grapevine virus A (GVA), the antiserum was successfully used in ELISA for the detection of GVB in grapevines.
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Porcine circovirus disease is currently the greatest threat to pig farming. Four main porcine circovirus genotypes are circulating worldwide.The study aimed to assess the prevalence of porcine circovirus genotypes in the central part of Shanxi province.We investigated the prevalence of porcine circovirus type 2 (PCV2), porcine circovirus type 3 (PCV3), and porcine circovirus type 4 (PCV4). Porcine circoviruses were analyzed by polymerase chain reaction (PCR) in the lung tissues of 180 pigs from 7 slaughterhouses in central Shanxi, China.The prevalence of PCV2, PCV3, and PCV4 were 56.8, 80, and 9.4%, respectively, and the negative rate was 10% for all three pathogens. The co-infection with PCV2 + PCV3, PCV2 + PCV4, PCV3 + PCV4, and PCV2 + PCV3 + PCV4 were 47.2, 7.4, 7.4, and 5.6%, respectively. Among PCV4-positive samples, the positive rate of PCV4 + PCV2 was 52.9% (9/17), whereas that of PCV4 + PCV3 was 100% (17/17). On the other hand, PCV2 and PCV3 were detected in 57.1% (93/163) and in 78.5% (128/163) of PCV4-negative samples, respectively. Phylogenetic analysis demonstrated that PCV2, PCV3, and PCV4 were not in the same clade and were distant from each other.The high positive rates of PCV3, PCV2 + PCV3, and PCV3 + PCV4 suggest that PCV3 may play a decisive role in PCV2 and PCV4 infections. Therefore, further control of PCV3 is needed to reduce the spread of the virus.
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We first report here the genome sequences of 4 rearranged porcine circovirus type 2 strains, JSTZ, ZJQDH1, ZJQDH2, and JSHM, isolated from porcine sera in China. The complete circular genomes of these isolates are 578, 483, 574, and 772 nucleotides in length, respectively. They are predicted to be defective interfering particles of porcine circovirus type 2. The findings will help us to understand molecular evolution of porcine circovirus type 2 and the relationship between porcine circovirus type 2 and diseases.
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Recombinant plasmids pcDNA3.1-orf2 expressing PCV2 capsid protein were transfected into P815 line cells mediated by liposomes.The positive anti-G418 monoclonal cells were screened and obtained in the medium with G418,and identified by PCR,RT-PCR and indirect immunofluorenscence assay.Subsequently,the stability of the positive cells expressing aimed protein was detected.The results showed that orf2 gene integrated into the genome of P815 cells,the capsid protein coded by PCV2 ORF2 was expressed in the acquired positive P815 monoclonal cells,and the P815 cell line with a stable expression of PCV2 capsid protein was obtained.The cell line will be necessary in the study of PCV2 new type vaccines,and the experiment method may be of referential value in other virus vaccines development.
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ABSTRACT We found a highly divergent circovirus in serum samples from several dogs. Phylogenetic analysis indicates that canine circovirus genotype 1 (CaCV-1) represents the first circovirus reported in dogs and is genetically most closely related to the only known mammalian circovirus, porcine circovirus. Here we report the complete genome sequence of the CaCV-1 strain NY214, which will help toward understanding the evolutionary and pathogenic characteristics of mammalian circoviruses.
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