title: Comparative analysis of the standard PCR-Based Replicon Typing (PBRT) with the commercial PBRT-KIT.
abstract: Plasmids are the main vectors of resistance and virulence genes in Enterobacteriaceae and plasmid typing is essential for the analysis of evolution, epidemiology and spread of antibacterial resistance. The PCR-Based Replicon Typing (PBRT), developed by Carattoli et al. in 2005, was an efficient method for plasmid identification and typing in Enterobacteriaceae. The 2005 PBRT scheme detected 18 replicons in 8 PCR reactions. Recently, the identification of novel replicons and plasmid types requested an update of the PBRT scheme. A commercial PBRT-KIT was devised for the identification of 28 different replicons in 8 multiplex PCRs. Here we report sensitivity and specificity of the PBRT-KIT carried out in comparison with the 2005 PBRT. The analysis of plasmid content was performed on forty-two enterobacterial strains from different sources, containing different replicon content. The 2005 PBRT identified replicons in 76.2% of the strains. The PBRT-KIT detected replicons in 100% of the analyzed strains, demonstrating increasing sensitivity and specificity of the commercial test with respect to the former 2005 PBRT scheme.
authors.affiliation: Department of Biomolecular Science, University of Urbino "Carlo Bo", via Arco d'Augusto, 2, 61032 Fano, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Carloni
authors.affiliation: Department of Biomolecular Science, University of Urbino "Carlo Bo", via Arco d'Augusto, 2, 61032 Fano, Italy. Electronic address: francesca.andreoni@uniurb.it.
authors.forename: Francesca
authors.initials: F
authors.lastname: Andreoni
authors.affiliation: Diatheva srl, via Sant'Anna, 131, 61030 Cartoceto, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Omiccioli
authors.affiliation: Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, viale Regina Elena, 299, 00161 Rome, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Villa
authors.affiliation: Department of Biomolecular Science, University of Urbino "Carlo Bo", via Arco d'Augusto, 2, 61032 Fano, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Magnani
authors.affiliation: Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, viale Regina Elena, 299, 00161 Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Carattoli
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004755:Enterobacteriaceae / Q000145:classification* / Q000235:genetics / Q000378:metabolism; D060885:Multiplex Polymerase Chain Reaction; D010957:Plasmids / Q000737:chemistry / Q000145:classification* / Q000378:metabolism; D011933:Reagent Kits, Diagnostic / Q000592:standards; D012093:Replicon; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic
keywords: Enterobacteriaceae; PCR-Based Replicon Typing; Plasmids; Resistance