Identification of Brucella ovis-specific proteins of 35 and 38 Kda
The immunoblotting technique has been used as an ancillary test in the diagnosis of Brucella ovis. The diagnosis is corroborated with complement fixation (CF), double agar gel immunodiffusion (DAGID), and enzyme-linked immunosorbent assay (ELISA). B. ovis hot saline (HS) extract has become the most popular antigen for these tests. Nevertheless, high degree of similarity has been observed between the protein bands from B. ovis and B. melitensis, resulting in serological cross reactions. This interferes with the diagnosis. The objective of this research was, therefore, identifying B. ovis-specific proteins and evaluating their antigenicity pattern by immunoblotting. B. ovis, B. melitensis, M. haemolytica, A. seminis, and H. somnus strains were used. Cellular fractions (outer membrane, inner membrane, and cytosol) from B. ovis Reo 198 strain, and B. melitensis 16M strain were obtained. Proteins were quantified, and then PAGE-SDS electrophoresis of both cellular fractions and whole extracts from all organisms was performed. Proteins were transferred to nitrocellulose membranes, and then treated with B. ovis hyperimmune serum. Three B. ovis-specific recognition bands (35, 38, and 81 kDa) -that were not found in B. melitensis or any of the other organisms- were identified. These proteins could be used in the diagnosis of B. ovis-caused epididymitis and, once properly standardized, they could have higher sensitivity/specificity for both DAGID and CF.
| Main Authors: | , , , |
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| Format: | Digital revista |
| Language: | spa |
| Published: |
Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias
2012
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| Online Access: | https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1411 |
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