Universal protocol for the rapid automated detection of carbapenem-resistant Gram-negative bacilli directly from blood cultures by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS)

Oviaño, Marina; Sparbier, Katrin; Barba, Maria José; Kostrzewa, Markus; Bou, Germán

doi:10.1016/j.ijantimicag.2016.08.024

Cited by 50 publications

(32 citation statements)

References 31 publications

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“…With the aim of establishing rapid and accurate procedures for detecting antimicrobial-resistant bacteria directly from positive blood cultures, Oviaño et al developed a universal method for detecting carbapenemase producers among 119 Gram-negative bacilli, including Enterobacteriaceae, Pseudomonas spp., and Acinetobacter spp., in 30 min, adding imipenem to a buffer containing NH 4 HCO 3 , ZnCl 2 , and sodium dodecyl sulfate (SDS) (66). NH 4 HCO 3 proved useful for detecting OXA-48 producers, as previously demonstrated by Papagiannitsis et al with meropenem (60).…”

Section: Detection Of ␤-Lactamasesmentioning

confidence: 82%

“…This field of work is progressing very rapidly, and since the last review, written by Hrabák et al (52) in 2013, some notable methodological advances have been made. (i) Incubation times have been reduced from around 3 h to 30 min by using ceftriaxone for ␤-lactamase detection (ESBL and AmpC) and by using imipenem for carbapenemase detection (55,61,66). (ii) A larger number of antibiotics have been tested, and ceftriaxone, cefpodoxime, and cefepime can now be analyzed (55).…”

Section: Detection Of ␤-Lactamasesmentioning

confidence: 99%

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Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

2018

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SUMMARY Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied in recent years for first-line identification of pathogens in clinical microbiology because it is simple to use, rapid, and accurate and has economic benefits in hospital management. The range of clinical applications of MALDI-TOF MS for bacterial isolates is increasing constantly, from species identification to the two most promising applications in the near future: detection of antimicrobial resistance and strain typing for epidemiological studies. The aim of this review is to outline the contribution of previous MALDI-TOF MS studies in relation to detection of antimicrobial resistance and to discuss potential future challenges in this field. Three main approaches are ready (or almost ready) for clinical use, including the detection of antibiotic modifications due to the enzymatic activity of bacteria, the detection of antimicrobial resistance by analysis of the peak patterns of bacteria or mass peak profiles, and the detection of resistance by semiquantification of bacterial growth in the presence of a given antibiotic. This review provides an expert guide for MALDI-TOF MS users to new approaches in the field of antimicrobial resistance detection, especially possible applications as a routine diagnostic tool in microbiology laboratories.

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Section: Detection Of ␤-Lactamasesmentioning

confidence: 82%

Section: Detection Of ␤-Lactamasesmentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

2018

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“…ammonium citrate, Tris-HCl, NH 4 HCO 3 ), different matrixes (i.e. HCCA and DHB) and/or different incubation times [2, 11, 20, 21, 22, 24, 25]. The increasing introduction of this assay in the clinical microbiology laboratories, also directly from positive blood cultures [25], has led to the development of a software for the automatic interpretation of the results, which can be helpful for nonexperienced users facing difficulties with spectrum analysis [21].…”

Section: Discussionmentioning

confidence: 99%

“…A normalised logRQ value comprised between 0.2 and 0.4 indicates an ambiguous hydrolysis measurement that requires re-testing or prolonging the incubation time [25]. Considering that our formula not completely overlaps that proposed by the manufacturer, in this study a cut-off for CAV was set at 0.3 on the basis of the results obtained from Panel II (91%, 131/144, with CAVs below 0.20 and only 9%, 13/144, with CAVs ranging from 0.21 to 0.25) and from carbapenemase-producing reference strains (CAVs ≥ 0.56).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a modified meropenem hydrolysis assay on a large cohort of KPC and VIM carbapenemase-producing Enterobacteriaceae

et al. 2017

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Carbapenem-resistant Enterobacteriaceae (CRE) have spread globally and represent a serious and growing threat to public health. The introduction of rapid and sensitive methods for the detection of carbapenemase-producing bacteria is of increasing importance. The carbapenemase production can be detected using non-molecular methods (such as the modified Hodge test, the synergy test, the Carba NP test and the antibiotic hydrolysis assays) and DNA-based methods. In this study, we propose a modified version of a previously described meropenem hydrolysis assay (MHA) by MALDI-TOF MS for the phenotypic detection in 2h of carbapenemase-producing Enterobacteriaceae. The MHA was successfully applied to detect carbapenemase activity in 981 well-characterized Enterobacteriaceae strains producing KPC or VIM carbapenemases, and in 146 carbapenem fully susceptible strains. This assay, applied also to NDM and OXA-48-producing strains and to CRE with resistance mechanisms other than carbapenemase production, has proved to be able to distinguish between carbapenemase-producing and -nonproducing Enterobacteriaceae.As already stated and as observed in our hands, MHA by MALDI-TOF MS analysis is independent from the type of carbapenemases involved, it is faster and easier to perform/interpret than culture-based methods. On the other hand, it cannot detect other carbapenem resistance mechanisms, such as porin alterations and efflux mechanisms.

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“…While most published protocols use pure culture, successful use of bacteria collected directly from positive blood cultures have also been reported. 50 While the protocol and analysis are yet to be standardized, use of MALDI-TOF MS appears to be a practical option for laboratories that have access to this technology.…”

Section: Phenotypic Testsmentioning

confidence: 99%

Carbapenem-Resistant Enterobacteriaceae

Iovleva

Doi

2017

Clinics in Laboratory Medicine

185

146

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Synopsis Carbapenem-resistant Enterobacteriaceae (CRE) have emerged as a major threat to modern medicine. Commonly used antibiotics are generally inactive against CRE. Therefore, timely detection of CRE in the clinical laboratory is of paramount importance. Among CRE, those producing carbapenem-hydrolyzing β-lactamase enzymes (carbapenemase-producing Enterobacteriaceae; CPE) are particularly of concern since they tend to spread among patients, and treatment of active infection is difficult. The carbapenemase groups most commonly encountered include KPC, NDM and OXA-48, with KPC overrepresented in the United States. Various phenotypic and genetic tests have been proposed and validated for rapid detection of CPE. Phenotypic tests detect carbapenemase activity either by inhibiting their activity with specific inhibitors of carbapenemases, or by detecting hydrolysis of carbapenems by carbapenemases. Genetic tests mostly depend on nucleic acid detection and amplification. Treatment options for CRE infections are limited and typically include combinations of polymyxins, tigecycline, aminoglycosides, or carbapenems, but newer agents with activity against CRE and better safety profiles are starting to become available and will likely emerge as the preferred therapy for the treatment of CRE infections in the near future.

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Universal protocol for the rapid automated detection of carbapenem-resistant Gram-negative bacilli directly from blood cultures by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS)

Cited by 50 publications

References 31 publications

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

Evaluation of a modified meropenem hydrolysis assay on a large cohort of KPC and VIM carbapenemase-producing Enterobacteriaceae

Carbapenem-Resistant Enterobacteriaceae

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