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Development and evaluation of an enzyme-linked immunosorbent assay for quantification of the humoral response of cattle vaccinated against Campylobacter fetus

María V. RepisoDirección de Laboratorios Veterinarios, Laboratorio Miguel C. Rubino, Ministerio de Ganadería, Agricultura y Pesca, Montevideo, Uruguay.

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Martín A. BaraibarDepartment of Immunology, Facultad de Química y Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.

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María A. OliveraDirección de Laboratorios Veterinarios, Laboratorio Miguel C. Rubino, Ministerio de Ganadería, Agricultura y Pesca, Montevideo, Uruguay.

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Silvia SilveyraDirección de Laboratorios Veterinarios, Laboratorio Miguel C. Rubino, Ministerio de Ganadería, Agricultura y Pesca, Montevideo, Uruguay.

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Julio BattistoniDepartment of Immunology, Facultad de Química y Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.

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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.

Animals—Twenty-six 24-month-old virgin Hereford heifers and a naturally infected Hereford bull.

Procedures—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 and Clinical Relevance—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. (Am J Vet Res 2002;63:586–590)

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.

Animals—Twenty-six 24-month-old virgin Hereford heifers and a naturally infected Hereford bull.

Procedures—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 and Clinical Relevance—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. (Am J Vet Res 2002;63:586–590)