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Clinical and Vaccine Immunology, June 2008, p. 911-915, Vol. 15, No. 6
1071-412X/08/$08.00+0     doi:10.1128/CVI.00046-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Use of the Brucella melitensis Native Hapten To Diagnose Brucellosis in Goats by a Rapid, Simple, and Specific Fluorescence Polarization Assay

Campo Experimental Río Bravo, Instituto Nacional de Investigaciones Forestales y Agrícolas y Pecuarias, Rio Bravo, Tam., México,¹ Laboratorio de Bacteriología, Centro Nacional de Investigaciones Disciplinarias en Microbiología, Instituto Nacional de Investigaciones Forestales y Agropecuarias, México D. F., México,² Universidad Autónoma de Nuevo León, Departamento de Inmunología y Virología, Facultad de Ciencias Biológicas, San Nicolás de los Garza, N. L., México,³ Campo Experimental General Terán, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, General Terán, N. L., México,⁴ Consorcio Técnico del Norte de México, Guadalupe Nuevo León, México,⁵ Campo Experimental Sur de Tamaulipas, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Altamira, Tamaulipas, México⁶

Received 5 February 2008/ Returned for modification 3 March 2008/ Accepted 17 March 2008

The performance of the fluorescence polarization assay (FPA) using the recently described Brucella melitensis native hapten and the Brucella abortus O-polysaccharide tracer was evaluated and compared with those of The World Organization for Animal Health tests related to indirect and competitive enzyme-linked immunosorbent assays as classification variables for goat sera obtained from a high-prevalence area where vaccination was performed; test series were also evaluated to increase the final specificity of the tests. Our results showed that the respective relative sensitivity and specificity were 99.7% and 32.5% for the rose Bengal test with a 3% cell concentration (RBT3), 92.8% and 68.8% for the rose Bengal test with 8% cell concentration (RBT8), 98.4% and 84.9% for the Canadian complement fixation test (CFT), 83.7% and 65.5% for the Mexican CFT, 98.4% and 81.0% for the buffered plate agglutination test (BPAT), and 78.1% and 89.3% for the fluorescence polarization assay (FPA). The use of the FPA as the secondary test significantly increased the final specificities of test combinations; the screening tests BPAT, RBT3, and RBT8 plus FPA resulted in 90%, 91.2%, and 91.3% final specificities, respectively, whereas for the combinations RBT3 plus Mexican CFT, RBT8 plus Mexican CFT, and BPAT plus Canadian CFT, the specificities were 65.5%, 63.2%, and 91.7%, respectively. The results suggested that the FPA may be routinely applied as an adaptable screening test for diagnosis of goat brucellosis, since its cutoff can be adjusted to improve its sensitivity or specificity, it is a rapid and simple test, it can be the test of choice when specificity is relevant or when an alternative confirmatory test is not available, and it is not affected by vaccination, thus reducing the number of goats wrongly slaughtered due to misdiagnosis.

Published ahead of print on 2 April 2008.