title: Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia.
abstract: Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease that is widespread in sub-Saharan Africa. It is caused by Mycoplasma mycoides subsp. mycoides, a bacterium belonging to the Mycoplasma mycoides cluster. In the absence of an efficient CBPP vaccine, improved and easy-to-use diagnostic assays for recurrent testing combined with isolation and treatment of positive animals represent an option for CBPP control in Africa. Here we describe the comprehensive screening of 17 immunogenic Mycoplasma mycoides subsp. mycoides proteins using well-characterized bovine sera for the development of a novel cocktail enzyme-linked immunosorbent assay (ELISA) for laboratory use. Two recombinant Mycoplasma immunogens, MSC_0136 and MSC_0636, were used to set up a standardized cocktail ELISA protocol. According to the results from more than 100 serum samples tested, the sensitivity and specificity of the novel cocktail ELISA were 85.6% and 96.4%, respectively, with an overall diagnostic accuracy comparable to that of the Office International des Epizooties (OIE)-prescribed serological assays. In addition, we provide a proof of principle for a field-applicable, easy-to-use commercially produced prototype lateral-flow test for rapid (<30-min) diagnosis of CBPP.
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Jena, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Heller
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya.
authors.forename: Nimmo
authors.initials: N
authors.lastname: Gicheru
authors.affiliation: Central Veterinary Laboratory, Windhoek, Namibia.
authors.forename: Georgina
authors.initials: G
authors.lastname: Tjipura-Zaire
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Muriuki
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya.
authors.forename: Mingyan
authors.initials: M
authors.lastname: Yu
authors.affiliation: National Institute of Agrarian and Veterinary Research, INIAV, IP, Animal Health Unit, Lisbon, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Botelho
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya.
authors.forename: Jan
authors.initials: J
authors.lastname: Naessens
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya.|Institute of Veterinary Bacteriology, University of Bern, Bern, Switzerland.
authors.forename: Joerg
authors.initials: J
authors.lastname: Jores
authors.affiliation: International Livestock Research Institute, Nairobi, Kenya a.liljander@cgiar.org.
authors.forename: Anne
authors.identifier: http://orcid.org/0000-0002-7543-5493
authors.initials: A
authors.lastname: Liljander
mesh_terms: D017954:Africa South of the Sahara; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D009176:Mycoplasma mycoides / Q000276:immunology*; D011002:Pleuropneumonia, Contagious / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000907:Antibodies, Bacterial