Sviluppo di metodiche molecolari per il rilevamento rapido di Pseudomonas in matrici alimentari e ambientali
Author(s)
Calisti, Cecilia
Date Issued
October 24, 2008
Type
Doctoral Thesis
Abstract
Main goal of this PhD project was to develop a rapid, efficient and sensitive procedures to identify and trace dissemination of pathogenic and spoilage Pseudomonas species in foods and environments. In this work, the molecular
identification of Pseudomonas species was achieved using new PCR-based assays with primer sets specific for 16S ribosomal DNA, rpoB and gyrB genes. These PCR assays were 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. Finally, we demonstrated that direct detection and identification of P. aeruginosa in environmental samples can be
achieved using a PCR technique based on the detection of gyrB. RFLP analysis of this latter gene can be also used for discriminating between P. aeruginosa and P. putida/P. fluorescens species.
Additional information
Dottorato di ricerca in Biotecnologie degli alimenti
File(s)![Thumbnail Image]()
Name
ccalisti_tesid.pdf
Size
5.69 MB
Format
Adobe PDF
Checksum (MD5)
e81bff0ea6f9801bf4571834da320a44