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The sera of the 25 S19-GFP vaccinated heifers (Table 1) were taken at 16 weeks after vaccination and used as positive controls. The negative control sera (n = 143) were taken from brucellosis-free unvaccinated controls (n = 118) and sera from all experimental heifers before vaccination (n = 25) and 118 Brucella infected bovines. The dotted arrow indicates the iELISA-GFP cut-off resulting in 100% diagnostic sensitivity and specificity was estimated as OD = 0.198.
