Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection
Abstract Background Porcine circovirus 2 (PCV-2) is one of the pathogens that leads to a growing and persistent threat in pigs. Thus, the development of serological detection methods for PCV-2 is of great necessity for clinical diagnosis as well as epidemiological investigations. This study aimed to...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-07-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-023-03648-7 |
| _version_ | 1827895122394611712 |
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| author | Yanwei Li Pingping Yu Yaxuan Bao Yuwen Ren Shaowei Zhao Xuexian Zhang |
| author_facet | Yanwei Li Pingping Yu Yaxuan Bao Yuwen Ren Shaowei Zhao Xuexian Zhang |
| author_sort | Yanwei Li |
| collection | DOAJ |
| description | Abstract Background Porcine circovirus 2 (PCV-2) is one of the pathogens that leads to a growing and persistent threat in pigs. Thus, the development of serological detection methods for PCV-2 is of great necessity for clinical diagnosis as well as epidemiological investigations. This study aimed to establish an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCV-2 based on virus-like particles (VLPs). Results Recombinant PCV-2 Cap protein was expressed in the baculovirus-insect cells system and PCV-2 VLPs were observed over transmission electron microscopy (TEM). The PCV-2 VLPs were shown to have good immunogenicity in mice and stimulated a high level of PCV-2 antibody titers. Using PCV-2 VLPs as coating antigen, the indirect ELISA can detect PCV-2 antibodies in animals with diagnostic sensitivity and specificity of 98.33% and 93.33% compared to immunofluorescence assay (IFA), respectively. The intra- and inter-assay coefficient variations (CVs) were < 10% in a batch, and < 15% in different batches, indicating good repeatability. There was no cross-reaction of this ELISA with antibodies against other porcine viruses. A total of 170 serum samples collected from different pig farms in China were tested for PCV-2 antibodies, and 151 (88.8%) samples were PCV-2 antibody positive. Conclusion Our findings suggest that this ELISA was rapid, specific, and reproducible and can be used for large-scale serological investigations of PCV-2 antibodies in pigs. |
| first_indexed | 2024-03-12T22:16:59Z |
| format | Article |
| id | doaj.art-86f21394a3174fb4a4f85683ef0a990c |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-03-12T22:16:59Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-86f21394a3174fb4a4f85683ef0a990c2023-07-23T11:17:25ZengBMCBMC Veterinary Research1746-61482023-07-011911910.1186/s12917-023-03648-7Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detectionYanwei Li0Pingping Yu1Yaxuan Bao2Yuwen Ren3Shaowei Zhao4Xuexian Zhang5Beijing Kemufeng Biopharmaceutical Co., LtdBeijing Kemufeng Biopharmaceutical Co., LtdBeijing Kemufeng Biopharmaceutical Co., LtdBeijing Kemufeng Biopharmaceutical Co., LtdBeijing Kemufeng Biopharmaceutical Co., LtdBeijing Kemufeng Biopharmaceutical Co., LtdAbstract Background Porcine circovirus 2 (PCV-2) is one of the pathogens that leads to a growing and persistent threat in pigs. Thus, the development of serological detection methods for PCV-2 is of great necessity for clinical diagnosis as well as epidemiological investigations. This study aimed to establish an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCV-2 based on virus-like particles (VLPs). Results Recombinant PCV-2 Cap protein was expressed in the baculovirus-insect cells system and PCV-2 VLPs were observed over transmission electron microscopy (TEM). The PCV-2 VLPs were shown to have good immunogenicity in mice and stimulated a high level of PCV-2 antibody titers. Using PCV-2 VLPs as coating antigen, the indirect ELISA can detect PCV-2 antibodies in animals with diagnostic sensitivity and specificity of 98.33% and 93.33% compared to immunofluorescence assay (IFA), respectively. The intra- and inter-assay coefficient variations (CVs) were < 10% in a batch, and < 15% in different batches, indicating good repeatability. There was no cross-reaction of this ELISA with antibodies against other porcine viruses. A total of 170 serum samples collected from different pig farms in China were tested for PCV-2 antibodies, and 151 (88.8%) samples were PCV-2 antibody positive. Conclusion Our findings suggest that this ELISA was rapid, specific, and reproducible and can be used for large-scale serological investigations of PCV-2 antibodies in pigs.https://doi.org/10.1186/s12917-023-03648-7Porcine circovirus 2Virus-like particlesIndirect ELISA |
| spellingShingle | Yanwei Li Pingping Yu Yaxuan Bao Yuwen Ren Shaowei Zhao Xuexian Zhang Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection BMC Veterinary Research Porcine circovirus 2 Virus-like particles Indirect ELISA |
| title | Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| title_full | Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| title_fullStr | Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| title_full_unstemmed | Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| title_short | Production of virus-like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| title_sort | production of virus like particles of porcine circovirus 2 in baculovirus expression system and its application for antibody detection |
| topic | Porcine circovirus 2 Virus-like particles Indirect ELISA |
| url | https://doi.org/10.1186/s12917-023-03648-7 |
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