Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus
Chicken parvovirus (ChPV) infection can cause runting-stunting syndrome (RSS) in chickens. There is currently no commercially available vaccine for controlling ChPV, and ChPV infection in chickens is widespread globally. The rapid detection of ChPV is crucial for promptly capturing epidemiological d...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-03-01
|
| Series: | Pathogens |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-0817/13/3/221 |
| _version_ | 1797239763784171520 |
|---|---|
| author | Minxiu Zhang Jianqi Liao Zhixun Xie Yanfang Zhang Sisi Luo Meng Li Liji Xie Qing Fan Tingting Zeng Jiaoling Huang Sheng Wang |
| author_facet | Minxiu Zhang Jianqi Liao Zhixun Xie Yanfang Zhang Sisi Luo Meng Li Liji Xie Qing Fan Tingting Zeng Jiaoling Huang Sheng Wang |
| author_sort | Minxiu Zhang |
| collection | DOAJ |
| description | Chicken parvovirus (ChPV) infection can cause runting-stunting syndrome (RSS) in chickens. There is currently no commercially available vaccine for controlling ChPV, and ChPV infection in chickens is widespread globally. The rapid detection of ChPV is crucial for promptly capturing epidemiological data on ChPV. Two monoclonal antibodies (mAbs), 1B12 and 2B2, against the ChPV NS1 protein were generated. A double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for detecting ChPV based on the mAb 1B12 and an anti-chicken polyclonal antibody against the ChPV NS1 protein. The detection limit for the ChPV recombinant pET32a-NS1 protein was approximately 31.2 ng/mL. A total of 192 throat and cloaca swab samples were analyzed for ChPV by the established DAS-ELISA and nested PCR methods. The concordance rate between the DAS-ELISA and the nested PCR method was 89.1%. The DAS-ELISA can detect the ChPV antigen without any cross-reaction with FAdV-4, FAdV-1, NDV, AIV, MS, CIAV, aMPV, EDSV, IBV, or AGV2. The method also has high repeatability, with a coefficient of variation (CV) of less than 5%. These findings indicate that the DAS-ELISA exhibits high accuracy, good sensitivity, and specificity, making it suitable for viral detection, field surveillance, and epidemiological studies. |
| first_indexed | 2024-04-24T17:56:43Z |
| format | Article |
| id | doaj.art-f438c8449e5a427695788c8342f8d4c0 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-04-24T17:56:43Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-f438c8449e5a427695788c8342f8d4c02024-03-27T13:58:52ZengMDPI AGPathogens2076-08172024-03-0113322110.3390/pathogens13030221Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken ParvovirusMinxiu Zhang0Jianqi Liao1Zhixun Xie2Yanfang Zhang3Sisi Luo4Meng Li5Liji Xie6Qing Fan7Tingting Zeng8Jiaoling Huang9Sheng Wang10Key Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaKey Laboratory of China (Guangxi)-ASEAN Cross-Border Animal Disease Prevention and Control, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, ChinaChicken parvovirus (ChPV) infection can cause runting-stunting syndrome (RSS) in chickens. There is currently no commercially available vaccine for controlling ChPV, and ChPV infection in chickens is widespread globally. The rapid detection of ChPV is crucial for promptly capturing epidemiological data on ChPV. Two monoclonal antibodies (mAbs), 1B12 and 2B2, against the ChPV NS1 protein were generated. A double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for detecting ChPV based on the mAb 1B12 and an anti-chicken polyclonal antibody against the ChPV NS1 protein. The detection limit for the ChPV recombinant pET32a-NS1 protein was approximately 31.2 ng/mL. A total of 192 throat and cloaca swab samples were analyzed for ChPV by the established DAS-ELISA and nested PCR methods. The concordance rate between the DAS-ELISA and the nested PCR method was 89.1%. The DAS-ELISA can detect the ChPV antigen without any cross-reaction with FAdV-4, FAdV-1, NDV, AIV, MS, CIAV, aMPV, EDSV, IBV, or AGV2. The method also has high repeatability, with a coefficient of variation (CV) of less than 5%. These findings indicate that the DAS-ELISA exhibits high accuracy, good sensitivity, and specificity, making it suitable for viral detection, field surveillance, and epidemiological studies.https://www.mdpi.com/2076-0817/13/3/221chicken parvovirusNS1 proteinmonoclonal antibodiesDAS-ELISA |
| spellingShingle | Minxiu Zhang Jianqi Liao Zhixun Xie Yanfang Zhang Sisi Luo Meng Li Liji Xie Qing Fan Tingting Zeng Jiaoling Huang Sheng Wang Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus Pathogens chicken parvovirus NS1 protein monoclonal antibodies DAS-ELISA |
| title | Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus |
| title_full | Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus |
| title_fullStr | Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus |
| title_full_unstemmed | Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus |
| title_short | Development of a Double-Antibody Sandwich ELISA Based on a Monoclonal Antibody against the Viral NS1 Protein for the Detection of Chicken Parvovirus |
| title_sort | development of a double antibody sandwich elisa based on a monoclonal antibody against the viral ns1 protein for the detection of chicken parvovirus |
| topic | chicken parvovirus NS1 protein monoclonal antibodies DAS-ELISA |
| url | https://www.mdpi.com/2076-0817/13/3/221 |
| work_keys_str_mv | AT minxiuzhang developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT jianqiliao developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT zhixunxie developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT yanfangzhang developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT sisiluo developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT mengli developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT lijixie developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT qingfan developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT tingtingzeng developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT jiaolinghuang developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus AT shengwang developmentofadoubleantibodysandwichelisabasedonamonoclonalantibodyagainsttheviralns1proteinforthedetectionofchickenparvovirus |