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

  • CECILIA CALISTI Dept. Agrobiology and Agrochemistry, University of Tuscia
  • MAURIZIO RUZII Via C. de Lellis, snc, I-01100 Viterbo, Italy
Keywords: Pseudomonas species, Pseudomonas aeruginosa, molecular typing, PCR, T-RFLP, 16S ribosomal DNA, gyrB gene.

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.

Author Biographies

CECILIA CALISTI, Dept. Agrobiology and Agrochemistry, University of Tuscia
ANIMAL SCIENCE
MAURIZIO RUZII, Via C. de Lellis, snc, I-01100 Viterbo, Italy
ANIMAL SCIENCE