Brucella is an expanding genus of Gram-negative intracellular wide host ranging pathogens. This work aimed at investigating molecular recognition of Brucella by MALDI-TOF MS as a rapid proteomic alternative to the bacteriologic gold standard. An MSP library of 11 reference Brucella strains including four species was created to cover the identification of the three classic Brucella species reported in Egypt.A dendrogram for reference strains was plotted to analyze proteomic relations. Based on bacteriologic and proteomic biotyping of 45 field isolates, a map revealed the geographic distribution of Brucella melitensis and B. abortus from 69 unvaccinated seropositive ruminants in 12 governorates during 2015. The MALDITOF MS was re-evaluated as a revolutionary molecular tool for Brucella identification reviewing the pros and cons of the technique suggesting recent methods to tackle existing hitches. It was concluded that bacteriologic and MALDI results fully matched thanks to the limited diversity of Brucella isolates and the narrow MSP library. For enhanced resolution
towards reliable distinction at the Brucella sub-species level, MALDI-TOF MS deserves selective enrichment of samples. Both B. melitensis Bv. 3 and B. abortus Bv. 1 were detected in cows and buffalo cows, while only B. melitensis Bv. 3 was recovered from small ruminants, a she camel and a man