title: Sensitive and specific enzyme-linked immunosorbent assay for detecting serum antibodies against Mycobacterium avium subsp. paratuberculosis in fallow deer.
abstract: The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies against Mycobacterium avium subsp. paratuberculosis in wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deer M. avium subsp. paratuberculosis isolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detect M. avium subsp. paratuberculosis antibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.
authors.affiliation: Department of Animal Health, Department of Agriculture of the Regional Government of the Principality of Asturias, SERIDA, Deva, Asturias, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Prieto
authors.affiliation: Department of Animal Health, Department of Agriculture of the Regional Government of the Principality of Asturias, SERIDA, Deva, Asturias, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Balseiro
authors.affiliation: Department of Animal Health, Department of Agriculture of the Regional Government of the Principality of Asturias, SERIDA, Deva, Asturias, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Casais
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Derio, Bizkaia, Spain.
authors.forename: Naiara
authors.initials: N
authors.lastname: Abendaño
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Derio, Bizkaia, Spain.
authors.forename: Liam E
authors.initials: LE
authors.lastname: Fitzgerald
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Derio, Bizkaia, Spain.
authors.forename: Joseba M
authors.initials: JM
authors.lastname: Garrido
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Derio, Bizkaia, Spain.
authors.forename: Ramon A
authors.initials: RA
authors.lastname: Juste
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Derio, Bizkaia, Spain malonso@neiker.net.
authors.forename: Marta
authors.initials: M
authors.lastname: Alonso-Hearn
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D003670:Deer / Q000097:blood / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D041981:Gastrointestinal Tract / Q000382:microbiology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology*; D016612:Paraffin Embedding; D010283:Paratuberculosis / Q000175:diagnosis* / Q000276:immunology; D012680:Sensitivity and Specificity; D014021:Tissue Preservation
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007074:Immunoglobulin G