Development and standardization of a western blotting test for detection of antibodies against B. abortus

Abstract

Brucellosis is an infectious disease caused by bacteria of the genus Brucella spp. with diagnosis based on use of serological techniques. The present study aimed to develop and standardize a western blotting (WB) test for detection of antibodies against B. abortus. Samples from two groups of cattle were analyzed: group I: 60 serum samples from true positive and true negative vaccinated animals (30 positive samples from infected animals according to rose bengal test (RBT), 2-mercaptoethanol, serum agglutination test (SAT) and complement fixation test (CFT) and 30 RBT negatives samples); group II: 383 field samples (90 positive and 293 CFT negative sera). The most reactive band in the western blotting, which properly identified and separated infected from non - infected had a molecular weight of ≤ 20kDa. The sensitivity, specificity and accuracy of the WB compared to RBT was 93%, 99%, 98%, respectively and k= 0.938. When compared to CFT, the sensitivity, specificity and accuracy of the WB was 97%, 98% and 97%, respectively and k= 0.929. The WB developed and standardized in the present study is a serological test with potential use as a confirmatory test for the diagnosis of bovine brucellosis.

A systematic review and meta-analysis of bovine brucellosis seroprevalence in Latin America and the Caribbean

“…After screening by abstract and title, 164 papers were selected for full-text assessment. Of these, 99 were excluded, and 65 studies [ [11] , [12] , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] , [31] , [32] , [33] , [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] , [42] , [43] , [44] , [45] , [46] , [47] , [48] , [49] , [50] , [51] , [52] , [53] , [54] , [55] , [56] , [57] , [58] , [59] , [60] , [61] , [62] , [63] , [64] , [65] , [66] , [67] , [68] , [69] , [70] , [71] , [72] , [73] , [74] , [75] ] were finally included for the final qualitative and quant...…”

“… 2019 Brazil Cattle 1,800 2 0.11 RBT [ 23 ] Falcão M et al. 2019 Brazil Cattle 383 94 24.54 RBT [ 34 ] Carvalho L et al. 2022 Brazil Cattle 964 2 0.21 RBT [ 56 ] Gädicke P et al.…”

Brucellosis detection and the role of Brucella spp. cell wall proteins

“…Falcão et al . [ 21 ] evaluated antibody responses in seropositive cattle with brucellosis and seronegative vaccinated animals, targeting protein bands in western blotting. The most antigenic band in the western blotting, which accurately distinguished seropositive from vaccinated cattle, had an MW of ≤20 kDa.…”