Thesis (M.Sc.) - Cairo University - Faculty of Veterinary Medicine - Department of Microbiology

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 proteomic fingerprinting as well as novel plastic antibodies by developing and characterizing molecularly-imprinted hydrogels to grasp all surface epitopes at one go for whole cell recognition of Brucellaabortus, B. melitensisand B. suis known to exist among livestock in Egypt. An MSP library of 11 reference Brucella strains was created. A dendrogram for reference strains was plotted to analyze phyloproteomic relations. Based on bacteriologic and proteomic biotyping of 45 field isolates, a map revealed the geographic distribution of Brucellamelitensis and B. abortus from 69 unvaccinated seropositive ruminants in 12 governorates during 2015.The MALDI-TOF 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. E{uFB00}ective bacterial recognition using a cell-imprinted polymer (CIP) formed on a 96-well microplate was achieved within 30 minutes.The polymer could discriminate the target strain from other strains with high selectivity reaching approximately 20 folds. It was concluded that bacteriologic and MALDI results fully matched thanks to the limited diversity of Brucella isolates and the narrow MSP library. The CIP approach proved valuable for rapid direct Brucella recognition and quantification colorimetrically in microtiter plates after full validation

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