Ghostbuster-A Western Blot-Based Panel Method to Resolve False-Positive Brucellosis Serology Test Results

False-positive serologic results (FPSRs) of brucellosis occur from time to time in various livestock with all the consequences (quarantine, compulsory slaughter, etc.) that follow true-positive laboratory results. A method based on the Polyacrylamide Gel Electrophoresis/Western Blot of a protein panel for resolving the FPSRs in the diagnosis of brucellosis was developed. Within the context of limited positive serum sample availability in Europe, the method successfully discriminates Brucella-positive sera from samples containing antibodies raised against infections caused by other Gram-negative bacteria causing FPSRs. An average CV% of 1.36 was determined for both repeatability and reproducibility for the whole separation mw range, and the test achieved 1.00 Diagnostic Sensitivity and 1.00 Diagnostic Specificity. The method with pre-prepared WB panels provides a rapid (less than 3 h), easily standardizable, and validatable alternative to existing confirmation methods. The whole WB process of the Brucella proteins and the subsequent densitometry can be accomplished with commercially available equipment, ready-to-use reagents, and open-source software, providing cost-effectiveness. The results of this study could attract broader attention, since molecular species in the 35.0-75.0 kDa range can serve as antigens in Brucella serology and the same fraction can be considered in the development of synthetic Brucella vaccines.