Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains

Abstract

Objective of this work was to develop the algorithms for differential PCR indication of Brucella genus strains using databases of their genomes. Materials and methods .Resources of the National Center for Biotechnology Information (NCBI) and BLAST and Vector NTI 9.1.0 software utilities. For PCR amplification, B. suis, B. abortus, B. melitensis nucleic acids, as well as plasmid DNA with marker insertions were used. Results and conclusions. We assessed brucella gene sequences, some of which are found in Brucella genus bacteria, others only in representatives of B. melitensis, and the third ones – only in representatives of B. abortus. As a result of primers and probes designing for indication of Brucella genus bacteria and representatives of B. melitensis and B. abortus species, criteria for marker sequence amplification have been established. These criteria provide for simultaneous differentiation in a single reaction. The determination of strain differences within one species of Brucella is described in multilocus VNTR assay technique, and the profiles of tandem repeats of various B. melitensis and B. abortus strains are available in the public domain. To monitor the progress of amplification, a positive control has been developed that has the nucleotide sequence of all marker regions. The text of the paper discloses all the nucleotide sequences of primers, probes and positive control, which makes it possible to independently acquire them in competent organizations.