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

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.