title: Development and evaluation of an enzyme-linked immunosorbent assay for quantification of the humoral response of cattle vaccinated against Campylobacter fetus.
abstract: OBJECTIVE
  To develop a reliable ELISA by use of a unique antigen preparation for serum IgG quantification after vaccination against Campylobacter fetus in cattle.
  
  
  METHODS
  Twenty-six 24-month-old virgin Hereford heifers and a naturally infected Hereford bull.
  
  
  METHODS
  5 antigens were prepared from a cell suspension of C fetus. Antigen preparations were the same as those reported in the literature, with the exception of antigens that were obtained by detergent solubilization of a C fetus cell suspension. For each antigen preparation, the optimal ELISA conditions for its immobilization were determined. Biotinylated antibodies against bovine immunoglobulins were obtained and used in the ELISA. Two groups of heifers were inoculated with commercial vaccines according to manufacturers' instructions. A control group was included. The immune response of vaccinated heifers and controls was followed for 6 months.
  
  
  RESULTS
  Detergent solubilized C fetus antigens resulted in better ELISA performance than other antigen preparations. Antigens were optimally immobilized at neutral pH and low ionic strength. All antigen preparations saturated the well with the same amount of protein. The vaccination schedule that advised a booster resulted in higher antibody titers, which were sustained over a longer period than the other schedule.
  
  
  CONCLUSIONS
  In the vaccination of cattle against C fetus, the ELISA we have developed may be used to evaluate serum antibody concentrations in response to various vaccines and vaccination schedules. Our results indicate that it is advisable to include a booster in the immunization protocol.
authors.affiliation: Dirección de Laboratorios Veterinarios, Laboratorio Miguel C. Rubino, Ministerno de Ganadería, Agricultura y Pesca, Montevideo, Uruguay.
authors.forename: Maria V
authors.initials: MV
authors.lastname: Repiso
authors.forename: Martin A
authors.initials: MA
authors.lastname: Baraibar
authors.forename: Maria A
authors.initials: MA
authors.lastname: Olivera
authors.forename: Silvia
authors.initials: S
authors.lastname: Silveyra
authors.forename: Julio
authors.initials: J
authors.lastname: Battistoni
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000276:immunology; D001428:Bacterial Vaccines / Q000276:immunology* / Q000592:standards; D002169:Campylobacter Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary*; D002168:Campylobacter fetus / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology* / Q000382:microbiology / Q000517:prevention & control; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D015151:Immunoblotting / Q000662:veterinary; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008297:Male; D014611:Vaccination / Q000379:methods / Q000662:veterinary*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; D007074:Immunoglobulin G