Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China
Abstract Background The existing detection methods for porcine circovirus type 2 (PCV2) specific antibodies in serum cannot determine the infection status, thus it is necessary to establish a method for detecting PCV2 antigen. The capsid protein (CAP) of PCV2, as a major structural protein that play...
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-022-03471-6 |
| _version_ | 1811336282811400192 |
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| author | Haojie Ding Yu Shen Yafan Gao Songrui Wu ChengZuo Xie Hao Sun Hongli Zhang Hongchao Sun Ying Shan Jianzu Ding Bin Zheng Shaohong Lu Xunhui Zhuo |
| author_facet | Haojie Ding Yu Shen Yafan Gao Songrui Wu ChengZuo Xie Hao Sun Hongli Zhang Hongchao Sun Ying Shan Jianzu Ding Bin Zheng Shaohong Lu Xunhui Zhuo |
| author_sort | Haojie Ding |
| collection | DOAJ |
| description | Abstract Background The existing detection methods for porcine circovirus type 2 (PCV2) specific antibodies in serum cannot determine the infection status, thus it is necessary to establish a method for detecting PCV2 antigen. The capsid protein (CAP) of PCV2, as a major structural protein that plays a significant role in viral replication and in inducing host’s immune response, is an ideal target antigen to monitor PCV2 infection. Therefore, a gold immunochromatographic assay (GICA) for rapid detection of PCV2 antigen based on the polyclonal antibodies (PAbs) against PCV2-CAP will be developed. Results The truncated CAP protein (dCAP) was used to immunize rabbits to generate anti-serum. After preliminary purification by caprylic acid/ammonium sulfate precipitation (CAAS), specific PAbs were purified by affinity chromatography column coupled with dCAP and its titer was about two-fold higher than preliminary purified PAbs. Colloidal gold-PAbs conjugate was synthesized under the optimum conditions. The specific anti-dCAP PAbs and goat anti-rabbit antibody (GAR) were then sprayed onto nitrocellulose (NC) membrane as a test line (TL) and a control line (CL), respectively. The visual limit detection (vLOD) of the GICA strips was 5 ng/mL. Specificity assay indicated that the GICA strips had specifically detected PCV2 and was not reactive for porcine epidemic diarrhea virus (PEDV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) or classic swine fever virus (CSFV). A total of 36 porcine serum samples were detected by this GICA and commercial enzyme-linked immunosorbent assay (ELISA) Kit, 9 positive samples were found by the developed strip with the rate of 25.0% comparing with 11 positive samples detected by the commercially ELISA Kit which positive rate was 30.5%, and the receiver operating characteristic (ROC) curve revealed that the relative sensitivity and specificity of this GICA strip were 72.7 and 96.0%, respectively, with an area of 87.2%. Conclusions This study established an efficient detection method with high sensitivity and specificity for the clinical diagnosis of PCV2 antigen, that will facilitate a rapid and convenient way to evaluate the infection status of vaccinated pigs. |
| first_indexed | 2024-04-13T17:37:11Z |
| format | Article |
| id | doaj.art-a64e1f64b2574113918a7473a8acf0cd |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-13T17:37:11Z |
| publishDate | 2022-10-01 |
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| series | BMC Veterinary Research |
| spelling | doaj.art-a64e1f64b2574113918a7473a8acf0cd2022-12-22T02:37:19ZengBMCBMC Veterinary Research1746-61482022-10-0118111110.1186/s12917-022-03471-6Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, ChinaHaojie Ding0Yu Shen1Yafan Gao2Songrui Wu3ChengZuo Xie4Hao Sun5Hongli Zhang6Hongchao Sun7Ying Shan8Jianzu Ding9Bin Zheng10Shaohong Lu11Xunhui Zhuo12School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeZhejiang Center of Animal Disease ControlDepartment of Animal Parasitology, Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural ScienceDepartment of Veterinary Medicine, College of Animal Sciences, Zhejiang UniversitySchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeSchool of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical CollegeAbstract Background The existing detection methods for porcine circovirus type 2 (PCV2) specific antibodies in serum cannot determine the infection status, thus it is necessary to establish a method for detecting PCV2 antigen. The capsid protein (CAP) of PCV2, as a major structural protein that plays a significant role in viral replication and in inducing host’s immune response, is an ideal target antigen to monitor PCV2 infection. Therefore, a gold immunochromatographic assay (GICA) for rapid detection of PCV2 antigen based on the polyclonal antibodies (PAbs) against PCV2-CAP will be developed. Results The truncated CAP protein (dCAP) was used to immunize rabbits to generate anti-serum. After preliminary purification by caprylic acid/ammonium sulfate precipitation (CAAS), specific PAbs were purified by affinity chromatography column coupled with dCAP and its titer was about two-fold higher than preliminary purified PAbs. Colloidal gold-PAbs conjugate was synthesized under the optimum conditions. The specific anti-dCAP PAbs and goat anti-rabbit antibody (GAR) were then sprayed onto nitrocellulose (NC) membrane as a test line (TL) and a control line (CL), respectively. The visual limit detection (vLOD) of the GICA strips was 5 ng/mL. Specificity assay indicated that the GICA strips had specifically detected PCV2 and was not reactive for porcine epidemic diarrhea virus (PEDV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) or classic swine fever virus (CSFV). A total of 36 porcine serum samples were detected by this GICA and commercial enzyme-linked immunosorbent assay (ELISA) Kit, 9 positive samples were found by the developed strip with the rate of 25.0% comparing with 11 positive samples detected by the commercially ELISA Kit which positive rate was 30.5%, and the receiver operating characteristic (ROC) curve revealed that the relative sensitivity and specificity of this GICA strip were 72.7 and 96.0%, respectively, with an area of 87.2%. Conclusions This study established an efficient detection method with high sensitivity and specificity for the clinical diagnosis of PCV2 antigen, that will facilitate a rapid and convenient way to evaluate the infection status of vaccinated pigs.https://doi.org/10.1186/s12917-022-03471-6Porcine circovirus type 2Capsid proteinGold immunochromatographic assayPolyclonal antibody |
| spellingShingle | Haojie Ding Yu Shen Yafan Gao Songrui Wu ChengZuo Xie Hao Sun Hongli Zhang Hongchao Sun Ying Shan Jianzu Ding Bin Zheng Shaohong Lu Xunhui Zhuo Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China BMC Veterinary Research Porcine circovirus type 2 Capsid protein Gold immunochromatographic assay Polyclonal antibody |
| title | Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China |
| title_full | Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China |
| title_fullStr | Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China |
| title_full_unstemmed | Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China |
| title_short | Development of gold Immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in Zhejiang province, China |
| title_sort | development of gold immunochromatographic assay strip based on specific polyclonal antibodies against capsid protein for rapid detection of porcine circovirus 2 in zhejiang province china |
| topic | Porcine circovirus type 2 Capsid protein Gold immunochromatographic assay Polyclonal antibody |
| url | https://doi.org/10.1186/s12917-022-03471-6 |
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