Differentiation of Antibodies against Selected Simbu Serogroup Viruses by a Glycoprotein Gc-Based Triplex ELISA
Abstract
:1. Introduction
2. Materials and Methods
2.1. Blood Samples
2.2. Production of Recombinant Proteins
2.2.1. Cloning
2.2.2. Expression and Purification in Drosophila S2 Cells
2.2.3. SDS-PAGE and Western Blot
2.3. ELISA Procedure
2.4. Data Analyses and Cut-off Determination
2.5. Repeatability and Reproducibility
3. Results
3.1. Expression and Purification of Proteins
3.2. Cut-off Determination and Diagnostic Characteristics
3.3. Repeatability and Reproducibility
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
References
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Animal Species | No. Negative | No. Positive against SBV | No. Positive against AKAV | No. Positive against SHUV |
---|---|---|---|---|
cattle | 192 | 76 | 30 | 42 |
sheep | 159 | 76 | 5 | 11 |
goat | 78 | 37 | 1 | 0 |
wild and zoo animals | 48 | 49 | 0 | 0 |
total | 477 | 238 | 36 | 53 |
Antigen | Animal Species | AUC | Sensitivity | Specificity |
---|---|---|---|---|
SBV | cattle-1 | 0.9519 (CI: 0.9259 to 0.9779) | 92.11% (CI: 83.60% to 97.05%) | 89.58% (CI: 84.37% to 93.52%) |
cattle-2 | 0.9418 (CI: 0.9149 to 0.9686) | 92.11% (CI: 83.60% to 97.05%) | 87.98% (CI: 83.10% to 91.86%) | |
sheep-1 | 0.9566 (CI: 0.9326 to 0.9806) | 93.42% (CI: 85.31% to 97.83%) | 86.79% (CI: 80.52% to 91.63%) | |
sheep-2 | 0.9532 (CI: 0.9289 to 0.9775) | 93.42% (CI: 85.31% to 97.83%) | 84.80% (CI: 78.52% to 89.82%) | |
goat-1 | 0.9186 (CI: 0.8553 to 0.9818) | 94.59% (CI: 81.81% to 99.34%) | 76.92% (CI: 66.00% to 85.71%) | |
goat-2 | 0.9196 (CI: 0.8571 to 0.9821) | 94.59% (CI: 81.81% to 99.34%) | 77.22% (CI: 66.40% to 85.90%) | |
wild/zoo animals-1 | 0.8835 (CI: 0.8199 to 0.9471) | 73.47% (CI: 58.92% to 85.05%) | 79.17% (CI: 65.01% to 89.53%) | |
wild/zoo animals-2 | 0.8835 (CI: 0.8199 to 0.9471) | 73.47% (CI: 58.92% to 85.05%) | 79.17% (CI: 65.01% to 89.53%) | |
overall-1 | 0.9339 (CI: 0.9159 to 0.9520) | 89.08% (CI: 84.40% to 92.74%) | 85.53% (CI: 82.05% to 88.57%) | |
overall-2 | 0.9292 (CI: 0.9110 to 0.9474) | 89.08% (CI: 84.40% to 92.74%) | 84.56% (CI: 81.20% to 87.53%) | |
AKAV | overall-1 | 0.9536 (CI: 0.9247 to 0.9825) | 69.44% (CI: 51.89% to 83.65%) | 97.06% (CI: 95.12% to 98.39%) |
overall-2 | 0.9386 (CI: 0.9079 to 0.9692) | 69.44% (CI: 51.89% to 83.65%) | 94.68% (CI: 92.80% to 96.19%) | |
SHUV | overall-1 | 0.9384 (CI: 0.8997 to 0.9771) | 84.91% (CI: 72.41% to 93.25%) | 93.29% (CI: 90.66% to 95.37%) |
overall-2 | 0.9234 (CI: 0.8792 to 0.9676) | 84.91% (CI: 72.41% to 93.25%) | 89.39% (CI: 86.89% to 91.55%) |
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Wernike, K.; Aebischer, A.; Sick, F.; Szillat, K.P.; Beer, M. Differentiation of Antibodies against Selected Simbu Serogroup Viruses by a Glycoprotein Gc-Based Triplex ELISA. Vet. Sci. 2021, 8, 12. https://doi.org/10.3390/vetsci8010012
Wernike K, Aebischer A, Sick F, Szillat KP, Beer M. Differentiation of Antibodies against Selected Simbu Serogroup Viruses by a Glycoprotein Gc-Based Triplex ELISA. Veterinary Sciences. 2021; 8(1):12. https://doi.org/10.3390/vetsci8010012
Chicago/Turabian StyleWernike, Kerstin, Andrea Aebischer, Franziska Sick, Kevin P. Szillat, and Martin Beer. 2021. "Differentiation of Antibodies against Selected Simbu Serogroup Viruses by a Glycoprotein Gc-Based Triplex ELISA" Veterinary Sciences 8, no. 1: 12. https://doi.org/10.3390/vetsci8010012