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Development and Evaluation of Epitope-Blocking ELISA for Detection of Antibodies against Contagious Caprine Pleuropneumonia in Goat Sera

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African Union-Pan African Veterinary Vaccine Centre (AU-PANVAC), P.O. Box 1746, Debrezeit, Ethiopia
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Molecular Biology and Biotechnology Department, Pan African University Institute for Basic Sciences, Technology and Innovation (PAUSTI), JKUAT Main Campus, P.O. Box 62000-00200, Nairobi, Kenya
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Biochemistry Department, Jomo Kenyatta University of Agriculture and Technology (JKUAT), P.O. Box 62000-00200, Nairobi, Kenya
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Research and Development Department, National Veterinary Institute, P.O. Box 19, Debrezeit, Ethiopia
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Department of Microbial Cellular and Molecular Biology, Addis Ababa University, P.O. Box 1176, Ethiopia
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Foot and Mouth Disease Department, Kenya Veterinary Vaccines Production Institute (KEVEVAPI), P.O. Box 53260-00200, Nairobi, Kenya
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Laboratoire Central Veterinaire, Laboratoire de Virologie, B.P. 206 Bingerville, Côte d’Ivoire
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Authors to whom correspondence should be addressed.
Vet. Sci. 2019, 6(4), 82; https://doi.org/10.3390/vetsci6040082
Received: 3 July 2019 / Revised: 6 September 2019 / Accepted: 7 September 2019 / Published: 18 October 2019
(This article belongs to the Section Microbiology and Immunology)
Enzyme linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies against contagious caprine pleuropneumonia (CCPP), the causative agent of which is Mycoplasma capricolum subsp. Capripneumoniae (Mccp). The currently available commercial CCPP competitive ELISA (CCPP cELISA) kit produced and supplied by IDEXX Company (Westbrook, Maine, United States) is relatively expensive for most African laboratories. To address this issue and provide a variety of choices, a sensitive and specific blocking-ELISA (b-ELISA) test to detect antibodies against CCPP was developed. We describe the newly developed CCPP blocking-ELISA based on the blocking of an epitope of a monoclonal antibody (Mccp-25) by a positive serum sample against the Mccp protein coated on a plate. The Percentage Inhibition (PI) cut-off value for the CCPP b-ELISA was set at 50 using 466 CCPP negative and 84 CCPP positive small ruminant sera. Of the negative sera, 307 were obtained from the Botswana National Veterinary Laboratory (BNVL) and 159 from the Friedrich-Loeffler-Institute (FLI) Germany. The 84 positive sera samples came from experimentally vaccinated goats at the AU-PANVAC facility in Debre-Zeit, Ethiopia. The relative diagnostic sensitivity and specificity of the CCPP b-ELISA was 93% and 88%, respectively. This test result indicated good correlation with that of the commercial CCPP cELISA by IDEXX Company (Westbrook, Maine, United States) with a Cohen’s κ agreement of κ agreement of 0.85. The newly developed CCPP b-ELISA will be useful in the detection of antibodies for the diagnosis CCPP and for sero-surveillance during vaccination campaigns. View Full-Text
Keywords: blocking-ELISA; cut-off value; monoclonal antibody; sensitivity and specificity blocking-ELISA; cut-off value; monoclonal antibody; sensitivity and specificity
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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.

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