DEVELOPMENT OF A MULTIPLEX PCR FOR IDENTIFICATION OF BRUCELLA SP. AND CROSS-REACTING YERSINIA ENTEROCOLITICA O:9

The aim of the study was to develop a multiplex PCR, which allows an identification of the universal 16S rRNA Brucella sp . marker and amplification of the perosamine synthet ase ( per ) gene, specific for cross-reacting Yersinia enterocolitica O:9 only. The PCR analysis of the DNA extracted from all Brucella reference strains used in the investigations reveal ed the presence of the 16S rRNA gene, generating the amplicon of 905 bp size. Para llely, the examination of the DNA from Y. enterocolitica belonging to O:9 serotype showed the presence of predicted ampli con of 312 bp typical to O:9 serotype only. The sen sitivity of the developed assay was determined with lymph tissue samples inoculated with B. abortus bv1 and Y. enterocolitica O:9 strains. The PCR analysis of the DNA extracted from lymph tissue revealed the presence of the predicted gene, when the 10 6 -10 2 CFU/g of the bacteria was added to the lymph tissue. The mPCR developed with direct extraction of DNA from lymph tissue can be a u seful tool for the differentiation of infections caused by Brucella and cross-reacting Y. enterocolitica O:9.

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