Evolution and Spread of a Multidrug-Resistant Proteus mirabilis Clone with Chromosomal AmpC-Type Cephalosporinases in Europe

Abstract

Proteus mirabilis isolates obtained in 1999 to 2008 from three European countries were analyzed; all carried chromosomal AmpC-type cephalosporinase bla CMY genes from a Citrobacter freundii origin (bla CMY-2 -like genes). Isolates from Poland harbored several bla CMY genes (bla CMY-4 , bla CMY-12 , bla CMY-14 , bla CMY-15 , and bla CMY-38 and the new gene bla CMY-45 ), while isolates from Italy and Greece harbored bla CMY-16 only. Earlier isolates with bla CMY-4 or bla CMY-12 , recovered in France from Greek and Algerian patients, were also studied. All isolates showed striking similarities. Their bla CMY genes resided within ISEcp1 transposition modules, named Tn6093, characterized by a 110-bp distance between ISEcp1 and bla CMY , and identical fragments of both C. freundii DNA and a ColE1-type plasmid backbone. Moreover, these modules were inserted into the same chromosomal site, within the pepQ gene. Since ColE1 plasmids carrying ISEcp1 with similar C. freundii DNA fragments (Tn6114) had been identified earlier, it is likely that a similar molecule had mediated at some stage this DNA transfer between C. freundii and P. mirabilis. In addition, isolates with bla CMY-12 , bla CMY-15 , and bla CMY-38 genes harbored a second bla CMY copy within a shorter ISEcp1 module (Tn6113), always inserted downstream of the ppiD gene. Sequence analysis of all mobile bla CMY-2 -like genes indicated that those integrated in the P. mirabilis chromosome form a distinct cluster that may have evolved by the stepwise accumulation of mutations. All of these observations, coupled to strain typing data, suggest that the bla CMY genes studied here may have originated from a single ISEcp1-mediated mobilization-transfer-integration process, followed by the spread and evolution of a P. mirabilis clone over time and a large geographic area.

Characterization of a Mobilizable IncQ Plasmid Encoding Cephalosporinase CMY-4 in Escherichia coli

“…1A). The right inverted repeat (IRR) of ISEcp1 was 116 bp upstream of bla , as in the CMY-encoding IncI and A/C plasmids (16). The bla CMY-4 -carrying region, however, lacked blc and sugE.…”

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Meropenem Hydrolysis Assay with NH 4 HCO 3 , a Reliable Tool for Direct Detection of Carbapenemase Activity

“…All isolates that were nonsusceptible to at least one carbapenem were subjected to metallo-␤-lactamase (M␤L), Klebsiella pneumoniae carbapenemase (KPC), and OXA-48 phenotypic detection using DDST with EDTA (10), the combined disk test with phenylboronic acid (11), and a temocillin disk (12), respectively. PCR detection of various bla genes was performed by the use of specific primers for bla SHV -, bla CTX-M -, bla CMY -, bla KPC -, bla VIM -, bla IMP -, bla NDM -, and bla OXA-48 -like genes, as reported previously (13)(14)(15)(16)(17)(18)(19)(20). The PCR products were purified and sequenced on both strands, as described previously (21).…”

Detection of CMY-99, a Novel Acquired AmpC-Type β-Lactamase, and VIM-1 in Proteus mirabilis Isolates in Bulgaria

“…The production of CMY-2 group variants by P. mirabilis isolates, which were found in four European countries, was reported by D'Andrea et al (2). Among them were isolates from Greece showing a combination of ␤-lactamases very similar to those of our isolates, namely, TEM-1, VIM-1, and CMY-16, which is the CMY variant closest related to CMY-99.…”