Abstract
Control of Malta fever caused by Brucella melitensis (B. melitensis) in humans depends on the reliability of diagnostic methods used for detection and identification of bacteria. In this study, we described a gold nanoparticles–DNA probes (AuNP–DNA probes) hybridization assay for detecting B. melitensis and its differentiation from other members of Brucella spp. in clinical samples. In this detection method, AuNP–DNA probes were hybridized in a tail-to-tail fashion with BMEII0466 sequence, which is specific to B. melitensis. When hybridization occurred in samples containing B. melitensis genomic DNA, it resulted in the formation of a complex and the solution color shifted from red to purple; therefore, visual detection is possible with the naked eye. The efficacy of AuNP–DNA probe hybridization assay was evaluated from genomic DNA of strains of B. melitensis and non-Brucella spp., and also genomic DNA from clinical samples. The results demonstrated that the main characteristics of this system include easy and rapid operation with high sensitivity and specificity. Therefore, no expensive techniques and equipments are necessary. This method can be employed as a regular diagnostic tool for B. melitensis in clinical samples.
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