title: Partition locus-based classification of selected plasmids in Klebsiella pneumoniae, Escherichia coli and Salmonella enterica spp.: an additional tool.
abstract: The dissemination of antimicrobial resistance genes in Enterobacteriaceae has been largely attributed to plasmids, circular DNA molecules capable of autonomous replication. Whereas high-copy-number plasmids primarily rely on passive diffusion for plasmid maintenance, low-copy-number plasmids utilize so-called partition (par) systems. Plasmid partition relies on three structures, i.e. a centromere like DNA site, a centromere-binding protein and an ATPase or a GTPase motor protein for plasmid positioning. Identification and classification of plasmids is essential for tracing plasmids conferring drug resistance. PCR-based replicon typing is currently the standard method for plasmid identification but there are new classification schemes, especially the relaxase gene typing (PRaseT). Here we developed a multiplex PCR set targeting par loci found on the plasmids most frequently encountered in Enterobacteriaceae. This method, called "plasmid partition gene typing" (PAR-T), was validated with 136 transconjugants or transformants harboring various replicon types. The method was tested with 30 multidrug-resistant clinical isolates including Escherichia coli, Klebsiella pneumoniae and Salmonella enterica subsp. enterica carrying 1-4 replicons; all replicons were tested in parallel with PRaseT for comparison. Six multiplex PCRs and one simplex PCR including 18 pairs of primers recognized plasmids of groups IncA/C, FIA, FIB, FIC, FIIk, FII, HI1, HI2, I1, L/M, N, X. Our set of multiplex PCRs showed high specificity for the classification of resistance plasmids except for IncX replicons.
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France. Electronic address: aurorebousquet@yahoo.fr.
authors.forename: A
authors.initials: A
authors.lastname: Bousquet
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Henquet
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France.
authors.forename: F
authors.initials: F
authors.lastname: Compain
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France.
authors.forename: N
authors.initials: N
authors.lastname: Genel
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France; AP-HP, Microbiology, Trousseau Hospital, Paris, France; AP-HP, Microbiology, Tenon Hospital, Paris, France; AP-HP, Microbiology, St-Antoine Hospital, Paris, France.
authors.forename: G
authors.initials: G
authors.lastname: Arlet
authors.affiliation: Sorbonne University, UPMC Univ Paris 06 CR7, Paris, France; INSERM U1135, CIMI, Team E13, Paris, France; AP-HP, Microbiology, St-Antoine Hospital, Paris, France.
authors.forename: D
authors.initials: D
authors.lastname: Decré
mesh_terms: D002503:Centromere / Q000235:genetics; D003198:Computer Simulation; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D007711:Klebsiella pneumoniae / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D010802:Phylogeny; D010957:Plasmids / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012093:Replicon; D019779:Salmonella enterica / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004247:DNA
keywords: Incompatibility group; Partition system; Plasmid