title: Use of an immunodominant P17 antigenic fraction of Neospora caninum in detection of antibody response in cattle.
abstract: A Neospora caninum 17 kDa protein fraction (p17) has been described as an immunodominant antigen (IDA) under reducing and non-reducing conditions. The aim of the present study was to investigate the diagnostic utility of p17 in cattle. In order to achieve this, p17 was purified by electroelution from whole N. caninum tachyzoite soluble extract and a p17-based Western blot (WB-p17) was developed. The p17 recognition was measured by densitometry and expressed as OD values to check the validity of the WB-p17. A total of 131 sera including sequential samples from naturally- and experimentally-infected calves and breeding cattle were analysed by WB-p17 and compared with IFAT using whole formalin-fixed tachyzoites as a reference test. The results obtained highlight the feasibility of using the N. caninum p17 in a diagnostic test in cattle. Firstly, the assay based on the p-17 antigen discriminated between known positive and negative sera from different cattle populations, breeding cattle and calves. Secondly, the p17 antigen detected fluctuations in the antibody levels and seroconversion in naturally- and experimentally-infected cattle. Significant differences in p-17 antigen recognition were observed between naturally infected aborting and non-aborting cattle, as well as significant antibody fluctuations over time in experimentally infected cattle, which varied between groups. Furthermore, the results obtained with WB-p17 are in accordance with the results obtained with the IFAT as high agreement values were obtained when all bovine subpopulations were included (kappa = 0.86).
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Avd. Puerta de Hierro s/n, 28040-Madrid, España
authors.forename: Gema
authors.initials: G
authors.lastname: Alvarez García
authors.forename: Inmaculada
authors.initials: I
authors.lastname: López Pérez
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Innes
authors.forename: Esther
authors.initials: E
authors.lastname: Collantes Fernandez
authors.forename: Aurora
authors.initials: A
authors.lastname: Fernandez Garcia
authors.forename: Mercedes
authors.initials: M
authors.lastname: Gomez Bautista
authors.forename: Luis Miguel
authors.initials: LM
authors.lastname: Ortega Mora
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D000953:Antigens, Protozoan / Q000276:immunology; D015153:Blotting, Western; D001947:Breeding; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000276:immunology; D003048:Coccidiosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D019084:Fluorescent Antibody Technique, Indirect; D016056:Immunodominant Epitopes / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D018688:Neospora / Q000276:immunology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G