Expanded-spectrum [beta]-lactamase and plasmid-mediated quinolone resistance.To the Editor: The emergence of plasmid-mediated, and thus transferable, quinolone resistance determinants has been recently discovered (1) and shown to involve the pentapeptide pen·ta·pep·tide
A polypeptide composed of five amino acids. repeat protein Qnr, which interacts with DNA gyrase DNA gyrase (ji´ras) a type II DNA topoisomerase. and topoisomerase IV to prevent quinolone inhibition (2,3). Qnr determinants confer resistance to nalidixic acid nalidixic acid /nal·i·dix·ic ac·id/ (nal-i-dik´sik) a synthetic antibacterial agent used in the treatment of genitourinary infections caused by gram-negative organisms.
n. and reduced susceptibility to fluoroquinolones (3). They have been identified worldwide in a variety of enterobacterial species and were often associated to expanded-spectrum [beta]3-lactamases (ESBLs) (2). The association between the ESBL ESBL Extended Spectrum Beta Lactamase
ESBL East Staffordshire Badminton League (UK) VEB-1 and the QnrA1 determinants was reported (4). Because plasmid co-localization of QnrA and VEB-1 encoding genes has been reported repeatedly from scattered clonally-unrelated enterobacterial isolates, our objective was to use replicon rep·li·con
A genetic element that undergoes replication as an autonomous unit. typing to trace a possible dissemination of a common plasmid worldwide.
The [bla.sub.VEB-1]- and/or qnrA-positive plasmids that have been included in the study were from 17 isolates previously described in detail (3-8) (Table). Escheriehia coli transconjugants (Tc) were obtained for 14 of 17 clinical isolates, allowing an accurate replicon typing since original clinical isolates might harbor several plasmids. They were collected from 1999 to 2005, from patients hospitalized in different parts of the world (Table). The 13 [bla.sub.VEB-1]-positive isolates were from 5 countries (France, Turkey, Algeria, Thailand, and Canada), scattered on 4 continents. Among them, the Providencia stuartii and Proteus mirabilis Proteus mirabilis Microbiology A gram-negative pathogen linked to UTIs, wound infections Habitat P mirabilis may be found in water,
soil, feces isolates from Algeria were negative for qnrA1. In addition, 4 [bla.sub.VEB-1]-negative but qnrA1-positive isolates recovered from France and Australia were also included in the study.
PCR-based replicon typing (PBRT), which recognizes FIA FIA
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To tell a fib. See Synonyms at lie2. , FIE fie
Used to express distaste or disapproval.
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FII Fundación Instituto de Ingeniería replicons (9), was applied to type the resistance plasmids from all the strains. Amplicons were confirmed by DNA sequencing and used as probes in hybridization hybridization /hy·brid·iza·tion/ (hi?brid-i-za´shun)
1. crossbreeding; the act or process of producing hybrids.
2. molecular hybridization
3. experiments on purified plasmids (data not shown).
PBRT results showed that the 13 [bla.sub.VEB-1]-positive plasmids (including 11 qnrAl-positive) belonged to the IncA/C incompatibility group incompatibility group Molecular biology A number of different types of plasmid, often related to each other, that are unable to stably coexist in the same cell . DNA sequencing identified the A/[C.sub.2] replicon variant (European Molecular Biology Laboratory The European Molecular Biology Laboratory (EMBL) is a molecular biology research institution supported by 19 countries comprising nearly all of western Europe and Israel. no. AM087198) in all these plasmids (Table). Plasmids of this type were recently identified in the United States and in Italy carrying the AmpC-type cephalosporinase CMY (Cyan Magenta Yellow) The color space used for printing. In theory, equal amounts of all three colors produce black. In practice, a separate black ink is required for quality printing. See CMYK. 2-encoding gene (10). In 2 strains (E. coli E. coli: see Escherichia coli.
in full Escherichia coli
Species of bacterium that inhabits the stomach and intestines. E. coli can be transmitted by water, milk, food, or flies and other insects. TcGOC and Citrobacter freundii Citrobacter freundii Microbiology A Citrobacter opportunistic pathogen Management Cephalothin, aminoglycosides LUT (LookUp Table) An array or matrix of values that contains data that is searched. See index and color palette. ), the IncA/[C.sub.2] plasmids were associated with additional replicons, which suggests the presence of multiple plasmids or fusions between plasmids of different backbones. By contrast, all the 4 [bla.sub.VEB-1]-negative isolates but qnrA1-positive were negative for the A/C replicon, except transconjugant TcK147; however, sequencing identified an A/[C.sub.1]-type replicon in that strain. These results indicated that the genes encoding QnrA1 and VEB-1, when identified concomitantly in a given isolate, were always located on plasmids belonging to the same IncA/[C.sub.2]-incompatibility group that may vary in size and digestion pattern (Table; unpub, data). In addition, we showed that plasmids carrying the [bla.sub.VEB-1] gene but lacking qnrA1 were also of the IncA/[C.sub.2] type (Table). Plasmids that were [bla.sub.VEB-1]-negative but qnrAl-positive were of distinct replicon types, thus suggesting independent acquisition of the qnrA1 gene on different plasmids. It is remarkable that since VEB-1 is apparently always encoded by IncA/[C.sub.2] plasmids, when genes for QnrA1 and VEB-1 are found together, they also occur on IncA/[C.sub.2] plasmids.
Thus, evidence here shows that the IncA/[C.sub.2] plasmid is the main vehicle of the [bla.sub.VEB-1] gene worldwide, on which the qnrA1 gene may be added. The possibility that both [bla.sub.VEB-1] and qnrA1 genes may be identified on a single genetic structure in several isolates has been recently shown with their identification within the same sull-type integron (6).
Because results of these experiments provided a good marker for tracing [bla.sub.VEB-1]-positive plasmids, and taking in account the property of A/C-type plasmids to have a broad range of hosts (note: this has not been demonstrated for the specific A/[C.sub.2] subgroup), we tried to amplify the A/[C.sub.2] replicon in a collection of 15 [bla.sub.VEB-1]-positive and clonally unrelated Pseudomonas aeruginosa Pseudomonas aeruginosa A normal soil inhabitant and human saprophyte that may contaminate various solutions in a hospital, causing opportunistic infection in weakened Pts Clinical Infective endocarditis in IVDAs, RTIs, UTIs, bacteremia, meningitis, 'malignant' isolates from France, Thailand, India, and Kuwait. The [bla.sub.VEB-1] gene was supposed to be chromosome-encoded in those isolates. PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) failed to give any positive results, confirming the absence of an IncA/C-type plasmid and also ruling out the hypothesis of IncA/[C.sub.2]-type plasmid co-integration at the origin of [bla.sub.VEB-1] acquisition in P. aeruginosa.
The spread of plasmids carrying a large array of resistance genes among Enterobacteriaceae is of concern since this provides a convenient genetic mechanism for a given strain to become panresistant to antimicrobial drugs. In particular, the recent identification of the Qnr determinants has shown that plasmids may provide resistance (or at least reduced susceptibility) to quinolones and fluoroquinolones, whereas they are already known to carry resistance to [beta]-lactams, aminoglycosides, chloramphenicol chloramphenicol (klōr'ămfĕn`əkŏl'), antibiotic effective against a wide range of gram-negative and gram-positive bacteria (see Gram's stain). It was originally isolated from a species of Streptomyces bacteria. , tetracycline tetracycline (tĕ'trəsī`klēn), any of a group of antibiotics produced by bacteria of the genus Streptomyces. They are effective against a wide range of Gram positive and Gram negative bacteria, interfering with protein , rifampin rifampin (rĭfăm`pĭn), antibiotic used in the treatment of tuberculosis. It is also used to eliminate the meningococcus microorganism from carriers and to treat leprosy, or Hansen's disease. , sulfonamides Sulfonamides Definition
Sulfonamides are medicines that prevent the growth of bacteria in the body.
Sulfonamides are used to treat many kinds of infections caused by bacteria and certain other microorganisms. , and disinfectants, pQR1 (4) or pl (6) are examples of well-characterized plasmids that mediate multidrug resistance multidrug resistance,
n the adaptation of tumor cells or infectious agents to resist chemotherapeutic agents. by carrying [bla.sub.VEB-1] and qnrA1, together with aminoglycoside aminoglycoside /ami·no·gly·co·side/ (-gli´ko-sid) any of a group of antibacterial antibiotics (e.g., streptomycin, gentamicin) derived from various species of Streptomyces resistance genes aadB, aacA1, and aadA1, chloramphenicol resistance gene cmlA, rifampin resistance gene arr2, disinfectant resistance gene qacI, and sulfonamides resistance gene sul1.
Our study showed that the IncA/ [C.sub.2]-type plasmids may be the source of such worldwide dissemination. It means that 1 plasmid scaffold has brought the same (or at least very similar) multidrug resistance to multiple enterobacterial species in different continents.
We thank S. Bernabeu for technical assistance.
This work was funded by grants from the European Community (6th PCRD PCRD Postgraduate Centre for Refugee Doctors (UK) , LSHM-CT-2005-018705).
(1.) Martinez-Martinez L, Pascual A, Jacoby GA. Quinolone resistance from a transferable plasmid. Lancet. 1998;351:797-9.
(2.) Robicsek A, Jacoby GA, Hooper DC. The worldwide emergence of plasmid-mediated quinolone resistance. Lancet Infect Dis. 2006;6:629-40.
(3.) Nordmann P, Poirel L. Emergence of plasmid-mediated resistance to quinolones in Enterobacteriaceae. J Antimicrob Chemother. 2005;56:463-9.
(4.) Poirel L, Van De Loo M, Mammeri H, Nordmann P. Association of plasmid-mediated quinolone resistance with extended-spectrum [beta]3-lactamase VEB-1. Antimicrob Agents Chemother. 2005;49:3091-4.
(5.) Girlich D, Poirel L, Leelaporn A, Karim A, Tribuddharat C, Fennewald M, et al. Molecular epidemiology molecular epidemiology Molecular medicine An evolving field that combines the tools of standard epidemiology–case studies, questionnaires and monitoring of exposure to external factors with the tools of molecular biology–eg, restriction endonucleases, of the integronlocated VEB-1 extended-spectrum [beta]3-lactamase in nosocomial nosocomial /noso·co·mi·al/ (nos?o-ko´me-il) pertaining to or originating in a hospital.
1. Of or relating to a hospital.
2. enterobacterial isolates in Bangkok, Thailand. J Clin Microbiol. 2001;39:175-82.
(6.) Poirel L, Pitout JD, Calvo L, Rodriguez-Martinez JM, Church D, Nordmann P. In vivo in vivo /in vi·vo/ (ve´vo) [L.] within the living body.
Within a living organism.
in vivo adv. selection of fluoroquinolone-resistant Escherichia coli isolates expressing plasmid-mediated quinolone resistance and expanded-spectrum [beta]3-lactamase. Antimicrob Agents Chemother. 2006;50:1525-7.
(7.) Rodriguez-Martinez JM, Poirel L, Pascual A, Nordmann P. Plasmid-mediated quinolone resistance in Australia. Microb Drug Resist. 2006; 12:99-102.
(8.) Poirel L, Leviandier C, Nordmann P. Prevalence and genetic analysis of plasmid-mediated quinolone resistance determinants QnrA and QnrS in Enterobacteriaceae in a French University Hospital. Antimicrob Agents Chemother. 2006;50:3992-7.
(9.) Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCR-based replicon typing. J Microbiol Methods. 2005;63:219-28.
(10.) Carattoli A, Miriagou V, Bertini A, Loli A, Colinon C, Villa L, et al. Replicon typing of plasmids encoding resistance to newer [beta]-lactams. Emerg Infect Dis. 2006; 12:1145-8.
Address for correspondence: Laurent Poirel, Service de Bacteriologie-Virologie, Hopital de Bicetre, 78 Rue du General Leclerc, 94275 Le Kremlin-Bicetre CEDEX, France; email: firstname.lastname@example.org
Laurent Poirel, * Laura Villa, ([dagger]) Alessia Bertini, ([dagger]) Johann D. Pitout, ([double dagger]) Patrice Nordmann, * and Alessandra Carattoli ([dagger])
* Hopital de Bicetre, le Kremlin-Bicetre, France; ([dagger]) Istituto Superiore di Sanita, Rome, Italy; and ([double dagger]) University of Calgary, Calgary, Alberta, Canada
Table. Features of the VEB-1- or QnrA-positive isolates used in this study * Year of Straint (Ref.) Sp. of origin Country isolation Escherichia coli E. coli Thailand 1999 TcE1 (5) E. coli TcE4 (5) E. coli Thailand 1999 E. coli TcE5 (5) E. coli Thailand 1999 E. coli TcE7 (5) E. coli Thailand 1999 E. coli TcE8 (5) E. coli Thailand 1999 E. coli TcEl6 (5) E. coli Thailand 1999 E. coli TcEl8 (5) E. coli Thailand 1999 E. coli Tc(pl) (5) E. coli Canada 2000 E. coli Tc(pQR1) E. coli France 2003 (4) E. coli Tc(GOC) (4) Enterobacter France 2003 cloacae Citrobacter freundii C. freundii Turkey 2004 LUT (3) Providencia stuartii P. stuartii Algeria 2004 15 (this study) E. coli TcMAA (this Proteus Algeria 2004 study) mirabilis E. coli TcK147 (7) Klebsiella Australia 2002 pneumoniae E. cloacae Al (8) E. cloacae France 2004 E. coli TcA2 (8) Enterobacter France 2005 aerogenes E. coli TcA3 (8) K. pneumoniae France 2005 Plasmid Straint (Ref.) size (kb) ESBL QnrA1 Escherichia coli 160 VEB-1 + TcE1 (5) E. coli TcE4 (5) 150 VEB-1 + E. coli TcE5 (5) 150 VEB-1 + E. coli TcE7 (5) 150 VEB-1 + E. coli TcE8 (5) 150 VEB-1 + E. coli TcEl6 (5) 140 VEB-1 + E. coli TcEl8 (5) 180 VEB-1 + E. coli Tc(pl) (5) 180 VEB-1 + E. coli Tc(pQR1) 180 VEB-1 + (4) E. coli Tc(GOC) (4) 190 VEB-1 + Citrobacter freundii ND VEB-1 + LUT (3) Providencia stuartii ND VEB-1 - 15 (this study) E. coli TcMAA (this 190 VEB-1 - study) E. coli TcK147 (7) 160 SHV-12 + E. cloacae Al (8) 75 SHV-12 + E. coli TcA2 (8) 150 SHV-12 + E. coli TcA3 (8) 40 - + Resistance Straint (Ref.) Replicon markers ([double dagger]) Escherichia coli A/[C.sub.2] NAL, K, SSS, C, TcE1 (5) RA E. coli TcE4 (5) A/[C.sub.2] NAL, K, TM, SSS, C, RA E. coli TcE5 (5) A/[C.sub.2] NAL, K, TM, SSS, SXT E. coli TcE7 (5) A/[C.sub.2] NAL, K, TM, SSS, C, RA E. coli TcE8 (5) A/[C.sub.2] NAL, K, TM, SSS, TE E, coli TcEl6 (5) A/[C.sub.2] NAL, K, TM, SSS, RA E, coli TcEl8 (5) A/[C.sub.2] NAL, K, SSS, C, RA E. coli Tc(pl) (5) A/[C.sub.2] NAL, K, SSS, C, RA E. coli Tc(pQR1) A/[C.sub.2] NAL, K, SSS, (4) C, RA, SXT E. coli Tc(GOC) (4) A/[C.sub.2], FIB NAL, K, TM, SSS, C Citrobacter freundii A/[C.sub.2], FIB, NA LUT (3) K Providencia stuartii A/[C.sub.2] NA 15 (this study) E. coli TcMAA (this A/[C.sub.2] K, TM, SSS, study) C, SXT E. coli TcK147 (7) HI2, A/[C.sub.1] NAL, K, TM, C, P TE, SXT E. cloacae Al (8) HI2 NA E. coli TcA2 (8) FII NAL, K, TM, TE E. coli TcA3 (8) 11, K NAL, K, TM, C, TE Ref., reference; ESBL, expanded-spectrum [beta]-lactamase; NAL, nalidixic acid; K, kanamycin; SSS, sulfonamides; C, chloramphenicol; RA, rifampin; TM, tobramycin; SXT, trimethoprim-sulfamethoxazole; TE, tetracycline; ND, not determinable; NA, not applicable. ([dagger]) indicates that this is a transconjugant or a transformant. ([double dagger]) Non-[beta]-lactam-associated markers.