A Rapid Antimicrobial Susceptibility Test for Klebsiella pneumoniae Using a Broth Micro-Dilution Combined with MALDI TOF MS

Wang, Gang; Song, Guobin; Xu, Yuanhong

doi:10.2147/idr.s305280

Cited by 12 publications

(8 citation statements)

References 32 publications

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“…Other rapid resistance detection methods, including the disappearance and appearance of characteristic antimicrobials hydrolysis peaks based on MALDI-TOF MS data, also demonstrated good results. 14 However, that approach requires additional culture time after bacterial isolation.…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection of Carbapenem-Resistant Klebsiella pneumoniae Using Machine Learning and MALDI-TOF MS Platform

Wang¹,

Xia²,

Wu³

et al. 2022

IDR

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Background Rapid detection of carbapenem-resistant Klebsiella pneumoniae (CRKP) is essential for specific antimicrobial therapy. Machine learning techniques combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can be used as a rapid, reliable, sensitive, and low-cost species identification method. Methods Clinically collected K. pneumoniae were subjected to MALDI-TOF MS analysis. A random forest (RF) algorithm and non-linear support vector machine (SVM) were used to construct the RF, SVM, and dimension reduction (SVM-K) models, and their performance was assessed for accuracy, sensitivity, specificity, and area under the subject worker curve (AUC). Results The RF, SVM and SVM-K models showed good classification performance with 0.88, 0.88, and 0.91 accuracy, 0.82, 0.85, and 0.89 sensitivity, 0.93, 0.92, and 0.94 specificity with an AUC of 0.9013, 0.9298, and 0.9356, respectively. For the SVM-K model, the optimal dimension reduction was 105 to 153, and the average accuracy was >0.9. The top 10 peak features of significance according to the RF algorithm with 6515 Da appeared in 56.8% of CRKP isolates and 5.3% of CSKP isolates, which indicated the best classification performance. Conclusion The three RF, SVM, and SVM-K models showed excellent classification performance differentiating the CRKP from CSKP; the SVM-K model was the best. Data analysis with machine learning combined with MALDI-TOF MS can be employed as a rapid and inexpensive alternative to existing detection methods.

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Section: Discussionmentioning

confidence: 99%

Rapid Detection of Carbapenem-Resistant Klebsiella pneumoniae Using Machine Learning and MALDI-TOF MS Platform

Wang¹,

Xia²,

Wu³

et al. 2022

IDR

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“…This study provides an idea for the detection of enzyme-producing AMR, and screening of resistant strains could be achieved by detecting the degradation of antibiotic substrates under the action of antimicrobial resistance enzymes. MALDI-TOF MS-based methods on detection of bacterial resistance are mainly divided into three categories: (1) detection of characteristic spectrum peaks of antimicrobial-resistant bacteria (Cordovana et al, 2018;Dortet et al, 2018;Li et al, 2019;Dortet et al, 2020); (2) detection of bacterial growth in the presence of antibiotics (Correa-Martıńez et al, 2019;Axelsson et al, 2020;Wang G. et al, 2021); and (3) detection of the antimicrobial agent change under the action of enzyme produced from antimicrobial resistant bacteria (Hoyos-Mallecot et al, 2014;Oviaño et al, 2014;Xie et al, 2019). The characteristic peak spectrum associated with AMR is the most ideal indicator to be detected by MALDI-TOF MS in principle.…”

Section: Discussionmentioning

confidence: 99%

“…However, the special medium limitations, background peak interference, and other factors have influenced the development of this method in clinical microbiology laboratories. The detection of bacterial growth in the environment with different antibiotic concentrations to obtain MIC is also an exploration direction through MALDI-TOF MS-based methods, but the MIC for antimicrobial-resistant bacteria may be lower than that for AST by the broth microdilution method (Wang G. et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

“…In this study, the detection principle adopted belongs to the third detection strategy as mentioned above. The huge potential of MALDI-TOF MS in the rapid detection of drug resistance has been demonstrated by the enzyme-mediated antibiotic hydrolysis assays (Wang G. et al, 2021). The premise of this method is that the mechanism of antimicrobial resistance in pathogens is inactivated by enzyme lysis/modification.…”

Section: Discussionmentioning

confidence: 99%

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Methodology Establishment and Application of VITEK Mass Spectrometry to Detect Carbapenemase-Producing Klebsiella pneumoniae

Lin

et al. 2022

Front. Cell. Infect. Microbiol.

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The ability of VITEK mass spectrometry (MS) in detection of bacterial resistance is currently under exploration and evaluation. In this study, we developed and validated a VITEK MS method to rapidly test carbapenemase-producing Klebsiella pneumoniae (CPKP). Solvents, antibiotic concentrations, crystal conditions and times, centrifugation speeds, and other factors were optimized to design a rapid sample pretreatment process for CPKP detection by VITEK MS. The related parameters of the mass spectrum were adjusted on the instrument to establish an CPKP detection mode. 133 clinically isolated strains of CPKP in the microbiology laboratory at the Shenzhen People’s Hospital from 2004 to 2017 were selected for accuracy evaluation. The fresh suspected strains from the microbiology laboratory in 2020 were used to complete the clinical verification. Two antibiotics, meropenem (MEM) and imipenem (IPM), were used as substrates. These two substrates were incubated with suspected CPKP, and the results were obtained by VITEK MS detection. Using this method, different types of CPKP showed different detection results and all the CPKP strains producing KPC-2 and IMP-4 carbapenemase were detected by VITEK MS. Thus, VITEK MS can be used for rapid detection of CPKP, especially for some common types of CPKP. This method provides high accuracy and speed of detection. Combined with its cost advantages, it can be intensely valuable in clinical microbiology laboratories after the standard operating procedures are determined.

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“…The prospect of the growth-based phenotypic RAST was partly hindered by the duration of the lag phase[28], which was the time needed for strains to cope with the constraints of their environment before proliferation. Within a population, the intensity of respiration repression [29] caused the considerable heterogeneity in the lag duration and growth rate between individuals. On the other hand, it was found that the main problem of the incubation with 100-fold dilution of BMD for a short time was the phenomenon of delayed phenotypic resistance.…”

Section: Discussionmentioning

confidence: 99%

Rapidly quantitative antimicrobial susceptibility testing for Klebsiella pneumoniae using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry under optimal conditions

Zhang

Chen

Dong

et al. 2021

Preprint

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Background Klebsiella pneumoniae infections, especially Carbapenem-resistant Klebsiella pneumoniae (CRKP), have become an “Urgent Threats” with high morbidity and mortality. Therefore, rapidly determining of the susceptibility and timely choosing an appropriate antibiotic were the important premises of the treatment of Klebsiella pneumoniae infections. The present study was first to explore the fitness of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) in quantitative rapid antimicrobial susceptibility testing (RAST) with optimal conditions. Methods Firstly, we optimized the methodology based on the formic acid extraction method. Taking broth micro-dilution method (BMD) as a reference method, we utilized 25-fold, 50-fold and 100-fold dilutions matching with appropriate incubation time to evaluate the consistency of minimum inhibition concentration (MIC) values derived from BMD and MALDI-TOF MS. Finally, the performances of optimal dilution and incubation time were verified through the reproducibility on different three days. Results Directly incubated in the EP tubes and skipped the washing step can not only simplify the procedure, but reduce the unnecessary loss of bacteria biomass to improve sensitivity. The optimal volumes of 70% formic acid/100% acetonitrile (FA/ACN) and supernatant on the target plate were 3 µL and 2 µL, respectively. Comparing the different combination, 25-fold dilution of the 0.5 McFarland suspension and 2hr incubation time resulted in best performance. The consistency between the MS-MIC and corresponding BMD-MIC values in CRKP and Carbapenem-susceptible Klebsiella pneumoniae (CSKP) strains accounted for 76.67% and 66.67%, respectively, which generated the substantial agreements between MS-MIC and BMD-MIC values for CRKP strains (Kappa=0.643) and the moderate agreements for CSKP strains (Kappa=0.476). Finally, the MALDI-TOF MS-based RAST showed a well repeatability. Conclusion Considerable potentials were demonstrated for universal and mechanism-indepent RAST by MALDI-TOF MS with optimal conditions, which strengthened its application in accelerating reporting time and clinical diagnosis.

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A Rapid Antimicrobial Susceptibility Test for Klebsiella pneumoniae Using a Broth Micro-Dilution Combined with MALDI TOF MS

Cited by 12 publications

References 32 publications

Rapid Detection of Carbapenem-Resistant Klebsiella pneumoniae Using Machine Learning and MALDI-TOF MS Platform

Rapid Detection of Carbapenem-Resistant Klebsiella pneumoniae Using Machine Learning and MALDI-TOF MS Platform

Methodology Establishment and Application of VITEK Mass Spectrometry to Detect Carbapenemase-Producing Klebsiella pneumoniae

Rapidly quantitative antimicrobial susceptibility testing for Klebsiella pneumoniae using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry under optimal conditions

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