Development of Innovative Molecular Methods for the Detection and the Identification of Pseudomonas spp. in Environmental and Clinical Samples

CECILIA CALISTI, MAURIZIO RUZII

Abstract


We have developed a rapid, reliable and sensitive method to analyze, detect and trace dissemination of pathogenic and spoilage Pseudomonas species in foods and environments. The molecular identification of Pseudomonas species was achieved using a PCR-based assay with primer sets specific for 16S ribosomal DNA and gyrB genes. This PCR assay was found to provide highly genus-specific detection and could be successfully used to identify Pseudomonas in microbial consortia where these bacteria were not abundant. By coupling the PCR assay for 16S rDNA gene to a T-RFLP technique, identification of Pseudomonas strains at species level could be obtained without cultivation. PCR with a combination of two target sequences (multiplex PCR) appeared to be the optimum choice for discriminating between Pseudomonas and closely related genera.

Keywords


Pseudomonas species; Pseudomonas aeruginosa; molecular typing; PCR; T-RFLP; 16S ribosomal DNA, gyrB gene.

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DOI: http://dx.doi.org/10.15835/buasvmcn-asb:66:1-2:3310




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