Development of an Indirect ELISA to Distinguish between Porcine Sapelovirus-Infected and -Vaccinated Animals Using the Viral Nonstructural Protein 3AB
Abstract
:1. Introduction
2. Materials and Methods
2.1. Serum Samples
2.2. Expression of Recombinant Protein
2.3. Purification of Recombinant Protein
2.4. Development of the Indirect ELISA Protocol
2.5. Determination of the ELISA Threshold Value
2.6. Positive Sample Sensitivity Assay
2.7. Assay Reproducibility
2.8. Assay Specificity
3. Results
3.1. Purification and Identification of the PSV 3AB Protein
3.2. Optimization of the Indirect ELISA Protocol
3.3. Determining the Threshold Value of the Established ELISA
3.4. Evaluation of the Diagnostic Reproducibility, Specificity of the Established ELISA Method, and Positive Serum Sensitivity
3.5. Detection in Clinical Samples
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Samples | Interbatch Variation | Intrabatch Variation | ||
---|---|---|---|---|
Mean + SD | CV (%) | Mean + SD | CV (%) | |
1 | 2.754 ± 0.014 | 0.51 | 2.732 ± 0.028 | 1.03 |
2 | 2.610 ± 0.006 | 0.22 | 2.639 ± 0.050 | 1.88 |
3 | 2.225 ± 0.039 | 1.75 | 2.240 ± 0.036 | 1.60 |
4 | 2.373 ± 0.050 | 2.10 | 2.388 ± 0.048 | 1.99 |
5 | 2.420 ± 0.043 | 1.76 | 2.430 ± 0.057 | 2.33 |
6 | 0.988 ± 0.011 | 1.10 | 1.025 ± 0.075 | 7.29 |
7 | 0.191 ± 0.009 | 4.61 | 0.189 ± 0.014 | 7.38 |
8 | 0.178 ± 0.004 | 1.98 | 0.167 ± 0.004 | 2.30 |
9 | 0.138 ± 0.002 | 1.20 | 0.128 ± 0.004 | 3.22 |
10 | 0.265 ± 0.003 | 1.13 | 0.272 ± 0.013 | 4.78 |
11 | 0.151 ± 0.005 | 3.28 | 0.152 ± 0.003 | 1.83 |
12 | 0.201 ± 0.002 | 1.06 | 0.200 ± 0.008 | 3.83 |
Samples | Positive | Negative | Concordance (%) | |
---|---|---|---|---|
Healthy | 46 | 0 | 46 | 100.0 |
Vaccinated | 75 | 0 | 75 | 100.0 |
Infected | 62 | 61 | 1 | 98.93 |
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Zhong, Z.; Li, B.; Tao, J.; Cheng, J.; Shi, Y.; Tang, P.; Jiao, J.; Liu, H. Development of an Indirect ELISA to Distinguish between Porcine Sapelovirus-Infected and -Vaccinated Animals Using the Viral Nonstructural Protein 3AB. Curr. Issues Mol. Biol. 2024, 46, 9821-9830. https://doi.org/10.3390/cimb46090583
Zhong Z, Li B, Tao J, Cheng J, Shi Y, Tang P, Jiao J, Liu H. Development of an Indirect ELISA to Distinguish between Porcine Sapelovirus-Infected and -Vaccinated Animals Using the Viral Nonstructural Protein 3AB. Current Issues in Molecular Biology. 2024; 46(9):9821-9830. https://doi.org/10.3390/cimb46090583
Chicago/Turabian StyleZhong, Zuchang, Benqiang Li, Jie Tao, Jinghua Cheng, Ying Shi, Pan Tang, Jiajie Jiao, and Huili Liu. 2024. "Development of an Indirect ELISA to Distinguish between Porcine Sapelovirus-Infected and -Vaccinated Animals Using the Viral Nonstructural Protein 3AB" Current Issues in Molecular Biology 46, no. 9: 9821-9830. https://doi.org/10.3390/cimb46090583