A Full MALDI-Based Approach to Detect Plasmid-Encoded KPC-Producing Klebsiella pneumoniae

Cordovana, Miriam; Kostrzewa, Markus; Glandorf, J.; Bienia, Michael; Ambretti, Simone; Pranada, Arthur B.

doi:10.3389/fmicb.2018.02854

Cited by 29 publications

(20 citation statements)

References 31 publications

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“…In detail, 131/139 (94.2 %) Klebsiella pneumoniae carbapenamase (KPC) - producing K. pneumoniae and 32/191 (16.7 %) methicillin-resistant Staphylococcus aureus (MRSA) were detected (results that were in accordance with the literature [20, 21]). Only 4/8 Bacteroides fragilis identified could be classified for the presence of the cfiA gene (all cfiA -negative).…”

Section: Resultssupporting

confidence: 84%

Rapid Sepsityper in clinical routine: 2 years’ successful experience

Cordovana

Zignoli

Ambretti

2020

Journal of Medical Microbiology

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Introduction. Rapid identification of the causative agent of sepsis is crucial for patient outcomes. Aim. The Sepsityper sample preparation method enables direct microbial identification of positive blood culture samples via matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS). Hypothesis/Gap statement. The implementation of the Sepsityper method in the routine practice could represent a fundamental tool to achieve a prompt identification of the causative agent of bloodstream infections, and therefore accelerate the adoption of the proper antibiotic treatment. Methodology. In this study, the novel rapid workflow of the MALDI Biotypr Sepsityper kit (Bruker Daltonik GmbH, Germany) was evaluated using routine samples from a 2-year period (n=6918), and dedicated optimized protocols for the microbial groups that were more difficult to identify were developed. Moreover, the use of the residual bacterial pellet to perform susceptibility testing using different methods (commercial broth microdilution, disc diffusion, gradient diffusion) was investigated. Results. The rapid Sepsityper protocol allowed the identification of 5470/6338 (86.3 %) monomicrobial samples at species level, with very good performance for all of the clinically most significant pathogens (2510/2592 enterobacteria, 631/669 Staphylococcus aureus and 223/246 enterococci were identified). Streptococcus pneumoniae , Bacteroides fragilis and yeasts were the most troublesome to identify, but the application of specific optimized protocols significantly improved their rate of identification (from 14.7–71.5 %, 47.8–89.7 % and 37.1–89.5 %, respectively). Specificity was 100 % (no identification was made for the false-positive samples). Further, the residual pellet proved to be suitable to investigate susceptibility to antimicrobials, enabling us to simplify the workflow and shorten the time to report. Conclusion. The Rapid Sepsityper workflow proved to be a reliable sample preparation method for identification and susceptibility testing directly from positive blood cultures, providing novel approaches for accelerated diagnostics of bloodstream infections.

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Section: Resultssupporting

confidence: 84%

Rapid Sepsityper in clinical routine: 2 years’ successful experience

Cordovana

Zignoli

Ambretti

2020

Journal of Medical Microbiology

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“…The principle of designed approach relies on the mass spectrometric evaluation of the carbapenem molecule after a short co-incubation with the bacterial strains, monitoring the distinct mass peaks of the hydrolyzed and non-hydrolyzed forms of the antibiotic resulting from carbapenemase-dependent β-lactam ring degradation [ 24 , 25 ]. Due to the low signal intensities of the typically observed hydrolyzed molecular forms (254 m/z), the internal standard provided with the MBT STAR Matrix is used as hydrolysis mass reference to calculate the hydrolyzation ratio.…”

Section: Resultsmentioning

confidence: 99%

Study on carbapenemase-producing bacteria by matrix-assisted laser desorption/ionization approach

et al. 2021

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The development of new techniques for the detection of carbapenemase activity is of great importance since the increased incident of resistance against carbapenems represents a serious threat to global public health. In this context, the matrix-assisted laser desorption/ionization approach already demonstrated to be a reliable tool for rapid carbapenemase detection. As a newly developed test, there is still a lack of in-depth analysis of its robustness and possible wider application. The main goal of this study was to evaluate the potential for using the design MBT STAR-Carba assay as the pre-characterization method for Enterobacterales and P. aeruginosa strains in terms of the produced classes of carbapenemases using modified procedure parameters—various suspension densities and incubation times. Moreover, its usefulness for the in-depth analysis and characterization of metallo-β-lactamases (MBL) was tested by applying inhibition assays. In this study, the designed assay proved to be a sensitive tool for the detection of carbapenemase hydrolytic activity, which can be successfully used to partially classify the class of carbapenemase present. Additionally, the use of defined high concentration suspensions would allow to shorten the incubation time to 1 minute for certain strains. Considering that the assay was also suitable to investigate the effect of different inhibitors on the MBL activity, it demonstrates far higher discriminatory potential than only a rapid routine carbapenemase detection tool and could be used as a susceptibility assay.

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“…Although MALDI-TOF MS was traditionally used for bacterial ID, recent novel applications of this approach suggest that it can be used to detect resistance markers such as β-lactamases by measuring bacterial growth in the presence of an antibiotic. 64,65 Although important advances have been made, MALDI-TOF MS recognizes only a limited number of resistance mechanisms and has a limited sensitivity, requiring more than 10 5 CFU/mL for accurate detection from clinical samples.…”

Section: Matrix-assisted Laser Desorption Ionization Time-of-flight Mmentioning

confidence: 99%

Novel Approaches to Hasten Detection of Pathogens and Antimicrobial Resistance in the Intensive Care Unit

Guillamet

Burnham

Kollef

2019

Semin Respir Crit Care Med

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Antibiotic resistance is recognized as a key determinant of outcome in patients with serious infections influencing empiric antibiotic practices especially for critically ill patients. Within the intensive care unit (ICU), nosocomial infections and increasingly community-onset infections are caused by multidrug-resistant bacteria. Escalating rates of antibiotic resistance adds substantially to the morbidity, mortality, and cost related to infections treated in the ICU. Both gram-positive organisms, such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci, and gram-negative bacteria, including Pseudomonas aeruginosa, Acinetobacter species, carbapenem-resistant Enterobacteriaceae, and extended spectrum β-lactamase producing organisms, are urgent threats. The rising rates of antimicrobial resistance have resulted in routine empiric administration of broad-spectrum antibiotics by clinicians to critically ill patients even when bacterial infection is microbiologically absent. Moreover, new broad-spectrum antibiotics are a challenge to use effectively while avoiding emergence of further resistance. Use of rapid diagnostic technologies (RDTs) will likely provide an important methodology for achieving this important balance. There is an urgent need for integrating the administration of new and existing antibiotics with RDTs in a way that is safe, cost-effective, applicable in all countries, and sustainable.

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A Full MALDI-Based Approach to Detect Plasmid-Encoded KPC-Producing Klebsiella pneumoniae

Cited by 29 publications

References 31 publications

Rapid Sepsityper in clinical routine: 2 years’ successful experience

Rapid Sepsityper in clinical routine: 2 years’ successful experience

Study on carbapenemase-producing bacteria by matrix-assisted laser desorption/ionization approach

Novel Approaches to Hasten Detection of Pathogens and Antimicrobial Resistance in the Intensive Care Unit

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