title: Identification of plasmids by PCR-based replicon typing.
abstract: The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Igamma, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.
authors.affiliation: Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. alecara@iss.it
authors.forename: Alessandra
authors.initials: A
authors.lastname: Carattoli
authors.forename: Alessia
authors.initials: A
authors.lastname: Bertini
authors.forename: Laura
authors.initials: L
authors.lastname: Villa
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Falbo
authors.forename: Katie L
authors.initials: KL
authors.lastname: Hopkins
authors.forename: E John
authors.initials: EJ
authors.lastname: Threlfall
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D004755:Enterobacteriaceae / Q000187:drug effects / Q000235:genetics; D004926:Escherichia coli / Q000145:classification / Q000187:drug effects / Q000235:genetics; D006801:Humans; D017720:Molecular Epidemiology; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012093:Replicon / Q000235:genetics; D019779:Salmonella enterica / Q000145:classification / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D012703:Serotyping
chemical_list: D004269:DNA, Bacterial