Development of a Highly Specific Monoclonal Antibody-Based Sandwich ELISA for Rapid Detection of Porcine Circovirus Type 3
Abstract
1. Introduction
2. Materials and Methods
2.1. Cells, Vectors, and Animals
2.2. Expression and Purification of PCV3 Cap Protein
2.3. Production of Anti-PCV3 Cap Antibodies
2.4. Characterization of Anti-PCV3 Cap mAb and pAb
2.5. Fine Mapping of Epitope Targeted by mAb
2.6. Optimization of DAS-ELISA Procedure
2.7. Performance Verification of DAS-ELISA
2.8. Statistical Analysis
3. Results
3.1. PCV3 Cap Antigen Preparation
3.2. Preparation and Characterization of mAb and pAb Against PCV3 Cap
3.3. Fine Mapping of Epitope Targeted by mAb Against PCV3 Cap
3.4. Optimal DAS-ELISA Procedure
3.5. Evaluation of DAS-ELISA
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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| Sample | OD450 | Mean Value | Standard Deviation | Coefficient of Variation | ||
|---|---|---|---|---|---|---|
| 1 | 1.354 | 1.435 | 1.377 | 1.389 | 0.0417 | 3.01% |
| 2 | 0.777 | 0.858 | 0.885 | 0.840 | 0.0562 | 6.69% |
| 3 | 0.269 | 0.265 | 0.254 | 0.263 | 0.0078 | 2.96% |
| 4 | 0.517 | 0.572 | 0.51 | 0.533 | 0.034 | 6.37% |
| Sample | OD450 | Mean Value | Standard Deviation | Coefficient of Variation | ||
|---|---|---|---|---|---|---|
| 1 | 1.393 | 1.331 | 1.317 | 1.347 | 0.0404 | 3.00% |
| 2 | 0.823 | 0.812 | 0.799 | 0.8113 | 0.012 | 1.48% |
| 3 | 0.21 | 0.206 | 0.215 | 0.2103 | 0.0045 | 2.14% |
| 4 | 0.591 | 0.58 | 0.647 | 0.606 | 0.0359 | 5.93% |
| DAS-ELISA | ||||||
|---|---|---|---|---|---|---|
| Positive | Negative | Total | Coincidence rate | Kappa value | ||
| qPCR | Positive | 51 | 3 | 54 | 93.33% | 0.837 |
| Negative | 2 | 19 | 21 | |||
| Total | 55 | 10 | 75 | |||
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Li, Z.; Zhao, J.; Tian, A.; Wu, H.; Chen, H.; Song, Y. Development of a Highly Specific Monoclonal Antibody-Based Sandwich ELISA for Rapid Detection of Porcine Circovirus Type 3. Viruses 2025, 17, 1340. https://doi.org/10.3390/v17101340
Li Z, Zhao J, Tian A, Wu H, Chen H, Song Y. Development of a Highly Specific Monoclonal Antibody-Based Sandwich ELISA for Rapid Detection of Porcine Circovirus Type 3. Viruses. 2025; 17(10):1340. https://doi.org/10.3390/v17101340
Chicago/Turabian StyleLi, Zhen, Jiaying Zhao, Ang Tian, Hao Wu, Huanchun Chen, and Yunfeng Song. 2025. "Development of a Highly Specific Monoclonal Antibody-Based Sandwich ELISA for Rapid Detection of Porcine Circovirus Type 3" Viruses 17, no. 10: 1340. https://doi.org/10.3390/v17101340
APA StyleLi, Z., Zhao, J., Tian, A., Wu, H., Chen, H., & Song, Y. (2025). Development of a Highly Specific Monoclonal Antibody-Based Sandwich ELISA for Rapid Detection of Porcine Circovirus Type 3. Viruses, 17(10), 1340. https://doi.org/10.3390/v17101340

