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ABSTRACT: Canine Brucellosis is a zoonotic disease caused by Brucella canis that affects canines’ reproductive system . The infection spreads rapidly in breeding kennels because of the presence of asymptomatic individuals. The most commonly used diagnostic methods are the blood culture and the 2ME-RSAT serological test. However, they show problems of sensitivity and specificity, so a Western Blot test could serve as a complement to obtain an accurate diagnosis of this infection. The aim of this work was to develop a Western Blot test for the detection of B. canis in dogs. Proteins of two strains of B. canis were used; M- and Brucella canis Oliveri strain. Sera from kennel dogs from the metropolitan area of the Aburrá Valley from the serum bank of the Biogenesis research group which were classified into 4 groups according to the 2ME-RSAT tests results and blood culture were used. SDS-Page was carried out at 12% and Western blot, using as primary antibody a mixture of 4 sera from each group at a 1:500 dilution, and as secondary antibody anti canine IgG peroxidase conjugate 1:6000 dilution, was used. Then the weight of the bands was analyzed and an immunoreactive band for each group was established. Differences in the immunoreactive bands for each analyzed group were observed. In group 2 (2ME-RSAT and positive blood culture) immunoreactive bands of approximately 70, 55, 48 and 40 kDa were observed. In group 3 (2ME-RSAT positive, blood culture negative) bands of approximately 55 and 40 kDa were observed. In group 4 (2ME-RSAT negative and blood culture positive) bands of 20, 18 and 12 kDa were observed. This Western Blot test allows to differentiate infected from uninfected dogs because of the ban profile for each group.
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References
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