Detection of AmpC β-lactamase-producing Gram-negative bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Liu, Cong; Ding, Shoufu; Huang, Ying; Wang, Zhongxin; Shen, Jilu; Ling, Helen; Xu, Yuanhong

doi:10.1016/j.jhin.2017.11.010

Cited by 9 publications

(8 citation statements)

References 18 publications

(17 reference statements)

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“…The first of the foresaid approaches, the enzymatic approach, employs MALDI-TOF MS to detect mass shifts as a result of hydrolysis or degradation in response to the activity of specific enzymes, such as β-lactamase , or carbapenemase. − Though this technique only requires 30–180 min of incubation prior to analysis in a commercially available MALDI-TOF MS, such an assay is very specific in its analysis and only works for detecting particular types of antibiotic resistance . In the proteomic approach, MS can also be used to quantitatively or semiquantitatively monitor the responses of pathogens in the presence of different antibiotics after 60–150 min of incubation. , The mass spectral profile of a microorganism incubated in only growth medium is used as an internal standard to calculate relative growth responses of the same microorganism subjected to antibiotics at the breakpoint concentration .…”

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confidence: 99%

“…103 A plethora of research has been performed using MS for rapid AST, including MALDI-TOF MS and laser ablation electrospray ionization MS. 104 Here, we will summarize two of the more common MALDI-TOF MS approaches, including detection of enzymatic activity and quantitative growth monitoring via proteomic analysis, both of which have been studied clinically to some extent. 105 The first of the foresaid approaches, the enzymatic approach, employs MALDI-TOF MS to detect mass shifts as a result of hydrolysis or degradation in response to the activity of specific enzymes, such as β-lactamase 106,107 or carbapenemase. 108−110 Though this technique only requires 30−180 min of incubation prior to analysis in a commercially available MALDI-TOF MS, such an assay is very specific in its analysis and only works for detecting particular types of antibiotic resistance.…”

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Recent Advances in the Race to Design a Rapid Diagnostic Test for Antimicrobial Resistance

et al. 2018

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Even with advances in antibiotic therapies, bacterial infections persistently plague society and have amounted to one of the most prevalent issues in healthcare today. Moreover, the improper and excessive administration of antibiotics has led to resistance of many pathogens to prescribed therapies, rendering such antibiotics ineffective against infections. While the identification and detection of bacteria in a patient's sample is critical for point-of-care diagnostics and in a clinical setting, the consequent determination of the correct antibiotic for a patienttailored therapy is equally crucial. As a result, many recent research efforts have been focused on the development of sensors and systems that correctly guide a physician to the best antibiotic to prescribe for an infection, which can in turn, significantly reduce the instances of antibiotic resistance and the evolution of bacteria "superbugs." This review details the advantages and shortcomings of the recent advances (focusing from 2016 and onward) made in the developments of antimicrobial susceptibility testing (AST) measurements. Detection of antibiotic resistance by genomic AST techniques relies on the prediction of antibiotic resistance via extracted bacterial DNA content, while phenotypic determinations typically track physiological changes in cells and/ or populations exposed to antibiotics. Regardless of the method used for AST, factors such as cost, scalability, and assay time need to be weighed into their design. With all of the expansive innovation in the field, which technology and sensing systems demonstrate the potential to detect antimicrobial resistance in a clinical setting?

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Recent Advances in the Race to Design a Rapid Diagnostic Test for Antimicrobial Resistance

et al. 2018

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“…A recent prospective study confirmed that the identification of the causative agent of sepsis by using MALDI TOF MS resulted in a shorter time frame for adequate antibiotic therapy. In this study, ampC-positive and Gram-negative sepsis patients rapidly identified using MALDI TOF MS received optimal treatment within 48 h [32].…”

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Current Scenario and Challenges in the Direct Identification of Microorganisms Using MALDI TOF MS

2021

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MALDI TOF MS-based microbial identification significantly lowers the operational costs because of minimal requirements of substrates and reagents for extraction. Therefore, it has been widely used in varied applications such as clinical, food, military, and ecological research. However, the MALDI TOF MS method is laced with many challenges including its limitation of the reference spectrum. This review briefly introduces the background of MALDI TOF MS technology, including sample preparation and workflow. We have primarily discussed the application of MALDI TOF MS in the identification of microorganisms. Furthermore, we have discussed the current trends for bioaerosol detection using MALDI TOF MS and the limitations and challenges involved, and finally the approaches to overcome these challenges.

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“…Before and after incubation at 37℃ overnight, the optical density (OD) value of each well was measured by a micro-plate reader (Biotek Synergy HT, USA) at 600 nm. The BMD-MIC values was considered as the lowest concentration of the IPM that completely inhibited the bacterial growth with the OD value increase ≤ 0.050 [33].…”

Section: Reference Resistance Determinationmentioning

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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Detection of AmpC β-lactamase-producing Gram-negative bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Abstract: This study demonstrated that MALDI-TOF MS can rapidly detect AmpC-producing strains.

Cited by 9 publications

References 18 publications

Recent Advances in the Race to Design a Rapid Diagnostic Test for Antimicrobial Resistance

Recent Advances in the Race to Design a Rapid Diagnostic Test for Antimicrobial Resistance

Current Scenario and Challenges in the Direct Identification of Microorganisms Using MALDI TOF MS

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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