Distribution of antibodies to porcine circovirus in swine populations of different breeding farms

Few studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm 3 . Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p<0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in studies of experimental and natural infection, and in monitoring the response to vaccination on commercial farms.INDEX TERMS: Enzyme-linked immunosorbent assay, isopycnic centrifugation, porcine circovirus type 2, sucrose cushion, antibody.

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“…CsCl gradient, because the PCV exhibits a fluctuation density in CsCl of approximately 1.37g/cm 3 (Tischer et al 1995).…”

Section: Correlation Between the Reference Test And Double-antibody Smentioning

confidence: 99%

A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection

et al. 2016

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“…However, in addition to domestic swine, PCV2 was also detected in wild boars (Sanchez et al, 2001;Ellis et al, 2003;Vicente et al, 2004;Cságola et al, 2006) and cattle (Nayar et al, 1999), and antibodies against the non-pathogenic viruses of the PCV1 group were also detected in cattle, mice and humans (Tischer et al, 1995). The presence of the virus in species other than swine is controversial, as according to another survey, PCV2 or antibodies to PCV2 were not detectable in cattle and sheep (Allan et al, 2000).…”

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confidence: 99%

Replication and transmission of porcine circovirus type 2 in mice

Cságola

Cadar

Tuboly

2008

Acta Veterinaria Hungarica

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Little information is known about infection, replication and transmission of porcine circovirus type 2 (PCV2) in species other than swine. Two sets of animal experiments were carried out to investigate the susceptibility of mice to PCV2 and to study their possible role in maintaining and transmitting the virus. In the first experiment 14 mice were inoculated with PCV2 by the intraperitoneal route with 5 × 10 2 TCID 50 of the PCV2-ROM strain (Cadar et al., 2007). In a second experiment 24 mice were divided into two groups (A and B); mice in Group A (n = 18) were inoculated orally with 1 × 10 5 TCID 50 PCV2-ROM and mice in Group B (n = 6) were left uninoculated until day 12 post inoculation (p.i.), when they were mixed with Group A. The animals were sacrificed at intervals for postmortem investigation and virus genome detection by polymerase chain reaction (PCR). The PCR results indicated that PCV2 could replicate in mice infected intraperitoneally or by the oral route, and that the virus can be transmitted directly from mouse to mouse.

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“…Two types of porcine circoviruses have been identified; PCV1 fails to induce disease in pigs following several experimental infections, whereas PCV2 is considered the primary aetiological agent that induces porcine circovirus-associated diseases, including postweaning multisystemic wasting syndrome (PMWS) (Tischer et al, 1995;Allan et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

The ORF3 protein of porcine circovirus type 2 promotes secretion of IL-6 and IL-8 in porcine epithelial cells by facilitating proteasomal degradation of regulator of G protein signalling 16 through physical interaction

Choi

Rho

Kim

et al. 2015

Journal of General Virology

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Porcine circovirus type 2 (PCV2) is the main aetiological agent of postweaning multisystemic wasting syndrome. The mechanism of pathogenicity associated with PCV2 infection is still not fully understood. Nevertheless, the fact that large amounts of proinflammatory cytokines within lymphoid tissues are released during the early stage of PCV2 infection may induce chronic inflammatory responses followed by the destruction of lymphoid tissues. However, how PCV2 infection causes an excessive inflammatory response in the host immune system during the early stage of PCV2 infection has still not been elucidated. In this study, we show that direct interaction between the PCV2 ORF3 and regulator of G protein signalling 16 (RGS16) within the cytoplasm of host cells leads to ubiquitin-mediated proteasomal degradation of RGS16. Facilitated degradation of the RGS16 by PCV2 ORF3 further enhances NFkB translocation into the nucleus through the ERK1/2 signalling pathway and increased IL-6 and IL-8 mRNA transcripts. Consequently, more severe inflammatory responses and leukocyte infiltration occur around host cells. This evidence may be the first clue explaining the molecular basis of how excessive amounts of proinflammatory cytokines within lymphoid tissues are released during the early stage of PCV2 infection.

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Distribution of antibodies to porcine circovirus in swine populations of different breeding farms

Cited by 81 publications

References 4 publications

A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection

A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection

Replication and transmission of porcine circovirus type 2 in mice

The ORF3 protein of porcine circovirus type 2 promotes secretion of IL-6 and IL-8 in porcine epithelial cells by facilitating proteasomal degradation of regulator of G protein signalling 16 through physical interaction

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