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

Abstract

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.

Development of Immunochromatographic Assay for the Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae Antibodies

“…The CFT initially used in the diagnosis of CCPP by MacOwan and Minette (1976) was more specific but less sensitive than the IHA test ( Samiullah, 2013 ). To improve the specificity and sensitivity, several enzyme-linked immunosorbent assay (ELISA) methods were developed to confirm the prevalence of CCPP ( Peyraud et al, 2014 ; Jean de Dieu et al, 2019 ). Although with high specificity, sensitivity, and suitability for large-scale testing, such as the PCR method, the ELISA test relied on specialized laboratory equipment resulting in the unfeasibility for field diagnosis.…”

“…It belongs to the Mycoplasma mycoides cluster, including five closely related species, subspecies, or biotypes ( Woubit et al, 2004 ). Mccp and the other pathogens in the M. mycoides cluster or peste des petits ruminants virus (PPRV) or pasteurellosis may induce similar respiratory symptoms in goats ( Jean de Dieu et al, 2019 ), so that Mccp infection cannot usually be diagnosed by physical examination alone ( Chota et al, 2019 ). In addition, challenges also exist in the culture isolation of the etiologic agent because of its specific medium requirements ( Teshome and Sori, 2021 ).…”

“…Although several methods have been conducted to the determination of CCPP ( Woubit et al, 2004 ; Lorenzon et al, 2008 ; Tharwat and Al-Sobayil, 2017 ; Lin et al, 2018 ; Jean de Dieu et al, 2019 ), they have certain drawbacks in either ease-of-use, sensitivity, specificity, cost, or dependence on specialized equipment or expertise. Immunochromatographic assay (ICA) is a powerful technique based on immunochromatographic procedure, which has been frequently used for the rapid detection of various biological specimens, especially specific antigens and antibodies of many diseases ( Liu et al, 2021 ).…”

Determination of Seroprevalence of Contagious Caprine Pleuropneumonia and Associated Risk Factors in Goats and Sheep Using Classification and Regression Tree

“…In fact, only 20 countries isolated Mccp because of the fastidious nature of the organism and the massive application of antibiotic treatment in suspected cases [ 18 ]. Therefore, serological tests were widely used to detect antibodies against Mccp such as complement fixation test, indirect hemagglutination, and latex agglutination [ 3 , 19 , 20 ]. Recently, a highly specific cELISA for CCPP has been developed and used on a large scale [ 21 , 22 ].…”

Alterations in acid-base balance, blood gases, and hematobiochemical profiles of whole-blood and thoracic fluid in goats with contagious caprine pleuropneumonia

“…Confirmation of Mccp is carried out by different methods, including microscopic examination of pulmonary exudation, histopathology of pulmonary parenchyma, molecular identification, and typing, namely, by polymerase chain reaction (PCR), detection of antigen in affected tissue by gel precipitin tests, and characterization by Available at www.veterinaryworld.org/Vol.14/July-2021/21.pdf modern serological techniques [1]. The more recently developed methodologies include the latex agglutination test (LAT), competitive enzyme-linked immunosorbent assay, PCR, and restriction fragment length polymorphism [1,[7][8][9]. Evaluation of cavity fluids including abdominal, thoracic, and pericardial effusions in the laboratory is beneficial for evaluating disease severity [10,11].…”