A set of multiplex PCRs for genotypic detection of extended-spectrum β-lactamases, carbapenemases, plasmid-mediated AmpC β-lactamases and OXA β-lactamases

Voets, Guido M.; Fluit, Ad C.; Scharringa, Jelle; Stuart, James Cohen; Hall, Maurine A. Leverstein–van

doi:10.1016/j.ijantimicag.2011.01.005

Cited by 90 publications

(57 citation statements)

References 14 publications

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“…Although it has not been tested directly on stool specimens, this NAAT showed 98% sensitivity and specificity for VIM-producing CPE when used on DNA extracted from clinical specimens, including blood, urine, pus, and respiratory samples, from Greece (172). Another multiplex endpoint PCR was developed by Voets et al and allows the detection of a wide range of resistance genes (173). Some of these multiplex assays were developed by independent laboratories and are not widely available to most clinical laboratories.…”

Section: Screening Methods To Detect Fecal Carriage Of Cposmentioning

confidence: 99%

Intestinal Carriage of Carbapenemase-Producing Organisms: Current Status of Surveillance Methods

et al. 2016

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SUMMARYCarbapenemases have become a significant mechanism for broad-spectrum β-lactam resistance inEnterobacteriaceaeand other Gram-negative bacteria such asPseudomonasandAcinetobacterspp. Intestinal carriage of carbapenemase-producing organisms (CPOs) is an important source of transmission. Isolation of carriers is one strategy that can be used to limit the spread of these bacteria. In this review, we critically examine the clinical performance, advantages, and disadvantages of methods available for the detection of intestinal carriage of CPOs. Culture-based methods (Centers for Disease Control and Prevention [CDC] protocols, chromogenic media, specialized agars, and double-disk synergy tests) for detecting carriage of CPOs are convenient due to their ready availability and low cost, but their limited sensitivity and long turnaround time may not always be optimal for infection control practices. Contemporary nucleic acid amplification techniques (NAATs) such as real-time PCR, hybridization assays, loop-mediated isothermal amplification (LAMP), or a combined culture and NAAT approach may provide fast results and/or added sensitivity and specificity compared with culture-based methods. Infection control practitioners and clinical microbiologists should be aware of the strengths and limitations of available methods to determine the most suitable approach for their medical facility to fit their infection control needs.

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Section: Screening Methods To Detect Fecal Carriage Of Cposmentioning

confidence: 99%

Intestinal Carriage of Carbapenemase-Producing Organisms: Current Status of Surveillance Methods

et al. 2016

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“…Simplex PCR assays targeting a single carbapenemase type have been used successfully in numerous studies, although there is no consensus regarding the oligonucleotide primers that should be used for each bla gene group. Multiplex and real-time PCR methods that allow the identification of multiple carbapenemase gene types and that further shorten the detection time, in the case of real-time PCR, have also been utilized (20,70,172,219,253). Also, real-time PCR assays can be followed by a melting curve step to allow the accurate identification of carbapenemase gene variants (163).…”

Section: Molecular Detection Of Carbapenemase Genesmentioning

confidence: 99%

Carbapenemases in Klebsiella pneumoniae and Other Enterobacteriaceae: an Evolving Crisis of Global Dimensions

et al. 2012

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SUMMARY The spread of Enterobacteriaceae , primarily Klebsiella pneumoniae , producing KPC, VIM, IMP, and NDM carbapenemases, is causing an unprecedented public health crisis. Carbapenemase-producing enterobacteria (CPE) infect mainly hospitalized patients but also have been spreading in long-term care facilities. Given their multidrug resistance, therapeutic options are limited and, as discussed here, should be reevaluated and optimized. Based on susceptibility data, colistin and tigecycline are commonly used to treat CPE infections. Nevertheless, a review of the literature revealed high failure rates in cases of monotherapy with these drugs, whilst monotherapy with either a carbapenem or an aminoglycoside appeared to be more effective. Combination therapies not including carbapenems were comparable to aminoglycoside and carbapenem monotherapies. Higher success rates have been achieved with carbapenem-containing combinations. Pharmacodynamic simulations and experimental infections indicate that modification of the current patterns of carbapenem use against CPE warrants further attention. Epidemiological data, though fragmentary in many countries, indicate CPE foci and transmission routes, to some extent, whilst also underlining the lack of international collaborative systems that could react promptly and effectively. Fortunately, there are sound studies showing successful containment of CPE by bundles of measures, among which the most important are active surveillance cultures, separation of carriers, and assignment of dedicated nursing staff.

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“…Fresh isolates were subcultured twice on 5% blood agar plates for 24 h at 35°C before MIC testing. A multiplex PCR assay with five different primer pairs was employed to detect genes encoding commonly acquired metallo-␤-lactamases (MBLs) (bla VIM , bla IMP , bla SIM , bla GIM , and bla SPM ) (10). The presence of genes encoding extended-spectrum beta-lactamases (ESBLs), plasmid-mediated AmpCs, NDM, and KPCs were determined using PCR (10).…”

Section: Bacterial Isolatesmentioning

confidence: 99%

In VitroActivity of Polymyxin B in Combination with Various Antibiotics against Extensively Drug-Resistant Enterobacter cloacae with Decreased Susceptibility to Polymyxin B

Cai

Lim

Teo

et al. 2016

Antimicrob Agents Chemother

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Against extensively drug-resistant (XDR) Enterobacter cloacae, combination antibiotic therapy may be the only option. We investigated the activity of various antibiotics in combination with polymyxin B using time-kill studies (TKS). TKS were conducted with four nonclonal XDR E. cloacae isolates with 5 log 10 CFU/ml bacteria against maximum, clinically achievable concentrations of polymyxin B alone and in two-drug combinations with 10 different antibiotics. A hollow-fiber infection model (HFIM) simulating clinically relevant polymyxin B and tigecycline dosing regimens was conducted for two isolates over 240 h. Emergence of resistance was quantified using antibiotic-containing (3؋ MIC) media. Biofitness and stability of resistant phenotypes were determined. All XDR E. cloacae isolates were resistant to all antibiotics except for polymyxin B (polymyxin B MIC, 1 to 4 mg/liter). All isolates harbored metallo-␤-lactamases (two with NDM-1, two with IMP-1). In single TKS, all antibiotics alone demonstrated regrowth at 24 h, except amikacin against two strains and polymyxin B and meropenem against one strain each. In combination TKS, only polymyxin B plus tigecycline was bactericidal against all four XDR E. cloacae isolates at 24 h. In HFIM, tigecycline and polymyxin B alone did not exhibit any killing activity. Bactericidal kill was observed at 24 h for both isolates for polymyxin B plus tigecycline; killing was sustained for one isolate but regrowth was observed for the second. Phenotypically stable resistant mutants with reduced in vitro growth rates were observed. Polymyxin B plus tigecycline is a promising combination against XDR E. cloacae. However, prolonged and indiscriminate use can result in resistance emergence.

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A set of multiplex PCRs for genotypic detection of extended-spectrum β-lactamases, carbapenemases, plasmid-mediated AmpC β-lactamases and OXA β-lactamases

Cited by 90 publications

References 14 publications

Intestinal Carriage of Carbapenemase-Producing Organisms: Current Status of Surveillance Methods

Intestinal Carriage of Carbapenemase-Producing Organisms: Current Status of Surveillance Methods

Carbapenemases in Klebsiella pneumoniae and Other Enterobacteriaceae: an Evolving Crisis of Global Dimensions

In VitroActivity of Polymyxin B in Combination with Various Antibiotics against Extensively Drug-Resistant Enterobacter cloacae with Decreased Susceptibility to Polymyxin B

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