Lab on a chip genotyping for Brucella spp. based on 15-loci multi locus VNTR analysis

Seventeen DNA samples of Brucella strains isolated in Sicily, previously genotyped, and twelve DNA samples, provided by MLVA Brucella VNTR ring trial, were analyzed by MLVA-15 on Agilent 2100. The DNA fragment sizes produced by Agilent, compared with those expected, showed discrepancies; therefore, in order to assign the correct alleles to the Agilent DNA fragment sizes, a conversion table was produced. In order to validate the system twelve unknown DNA samples were analyzed by this method obtaining a full concordance with the VNTR ring trial results.In this paper we described a rapid and specific detection method for the characterization of Brucella isolates. The comparison of the MLVA typing data produced by Agilent system with the data obtained by standard sequencing or ethidium bromide slab gel electrophoresis showed a general concordance of the results. Therefore this platform represents a fair compromise among costs, speed and specificity compared to any conventional molecular typing technique.The genus Brucella comprises Gram-negative bacteria that are the etiological agents of brucellosis, a zoonosis that represents one of the most important worldwide disease affecting animals and humans, especially in the Mediterranean areas [1,2]. The disease can be transmitted to humans directly by contact with infected animals or indirectly by contaminated dairy products. Brucella infections can cause chronic debilitating diseases in humans with the involvement of multiple organs and low mortality while in the domesticated animals can lead to reproductive failure with subsequent economic loss. Six classically species are recognized within the genus Brucella: Brucella abortus (7 biovars), Brucella melitensis (3 biovars), Brucella suis (5 biovars), Brucella ovis, Brucella canis, and Brucella neotomae [3]. Isolation and characterization of novel different Brucella strains from a wide variety wildlife species from terrestrial and marine mammals, recently classified as three