Development and Evaluation of Epitope-Blocking ELISA for Detection of Antibodies against Contagious Caprine Pleuropneumonia in Goat Sera
Abstract
:1. Introduction
2. Material and Methods
2.1. Ethical Approval
2.2. Preparation of Mycoplasma Capricolum Subsp. Capripneumoniae (Mccp) Antigen
2.3. Serum Samples
2.4. Monoclonal Antibody Production, Its Conjugation, and Evaluation in Epitope Blocking
2.5. CCPP Blocking-ELISA Development and Procedure
2.6. Evaluation of CCPP Blocking ELISA
2.6.1. Analytical Sensitivity
2.6.2. Cut-Off Value Determination
2.6.3. Assay Repeatability
2.6.4. Determination of Diagnostic Sensitivity and Specificity
3. Results
3.1. Titration of Mccp Antigen and Conjugate Used in the CCPP b-ELISA
3.2. Cut-Off Value Determination
3.3. Analytical Sensitivity and Blocking Epitope by Serum Samples
3.4. Assay Repeatability
3.5. Comparison of the Newly Developed CCPP b-ELISA with the Available Commercial CCPP cELISA
3.6. Diagnostic Sensitivity and Specificity
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Type of Test | Tested (+) | Tested (−) |
---|---|---|
Test (+) | T+ | F− |
Test (−) | F+ | T− |
Sera | Commercial CCPP cELISA | CCPP b-ELISA | ||
---|---|---|---|---|
Positive ≥ 55 PI | Negative < 55 PI | Positive ≥ 50 PI | Negative < 50 PI | |
252 | 90 | 162 | 104 | 148 |
Newly Developed Test | Commercial CCPP cELISA | Total | ||
---|---|---|---|---|
Positive | Negative | |||
CCPP b-ELISA | Positive | 84 | 20 | 104 |
Negative | 6 | 142 | 148 | |
Total | 90 | 162 | 252 |
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Share and Cite
Jean de Dieu, B.; Charles, B.S.; Nwankpa, N.; Chitsungo, E.; Moustapha Boukary, C.R.; Maina, N.; Tefera, T.A.; Nwankpa, R.V.; Mwangi, N.; Mathurin Koffi, Y. Development and Evaluation of Epitope-Blocking ELISA for Detection of Antibodies against Contagious Caprine Pleuropneumonia in Goat Sera. Vet. Sci. 2019, 6, 82. https://doi.org/10.3390/vetsci6040082
Jean de Dieu B, Charles BS, Nwankpa N, Chitsungo E, Moustapha Boukary CR, Maina N, Tefera TA, Nwankpa RV, Mwangi N, Mathurin Koffi Y. Development and Evaluation of Epitope-Blocking ELISA for Detection of Antibodies against Contagious Caprine Pleuropneumonia in Goat Sera. Veterinary Sciences. 2019; 6(4):82. https://doi.org/10.3390/vetsci6040082
Chicago/Turabian StyleJean de Dieu, Baziki, Bodjo S. Charles, Nick Nwankpa, Ethel Chitsungo, Cisse Rahamatou Moustapha Boukary, Naomi Maina, Takele A. Tefera, Rume Veronica Nwankpa, Nduta Mwangi, and Yao Mathurin Koffi. 2019. "Development and Evaluation of Epitope-Blocking ELISA for Detection of Antibodies against Contagious Caprine Pleuropneumonia in Goat Sera" Veterinary Sciences 6, no. 4: 82. https://doi.org/10.3390/vetsci6040082