Identification of plasmids by PCR-based replicon typing.
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.
Carattoli
Alessandra
A
Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. alecara@iss.it
Bertini
Alessia
A
Villa
Laura
L
Falbo
Vincenzo
V
Hopkins
Katie L
KL
Threlfall
E John
EJ
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
D004269:DNA, Bacterial