MICy: a Novel Flow Cytometric Method for Rapid Determination of Minimal Inhibitory Concentration

Kállai, András; Kelemen, Márta; Molnár, Noémi; Tropotei, Adrienn; Hauser, Balázs; Iványi, Zsolt; Gál, János; Ligeti, Erzsébet; Kristóf, Katalin; Lörincz, Ákos M.

doi:10.1128/spectrum.00901-21

Cited by 7 publications

(3 citation statements)

References 32 publications

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“…However, they take several days to provide a result because they require the isolation of bacteria isolated from patients who were then exposed to the antibiotics. To overcome this delay in conventional AST assays, new phenotypic strategies have been proposed for the rapid measurement of various phenotypic characteristics of bacteria, such as morphology, metabolism, biochemical composition, and growth after exposure to antibiotics, including isothermal microcalorimetry [ 28 , 29 ], electrochemical impedance spectroscopy [ 30 , 31 ], microscopy [ 32 , 33 ], electrochemical ASTs [ 34 , 35 ], spectroscopy [ 36 ], flow cytometry [ 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 ], and spectrometry [ 46 ]. Rapid phenotypic AST methods have detected microbial growth and/or metabolism as well as morphological changes in biological samples with very low microbial loads.…”

Section: The Landscape Of Rapid Methods For Antibiotic Susceptibility...mentioning

confidence: 99%

Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Măruţescu

2023

Microorganisms

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Antimicrobial resistance is a global threat to human health and welfare, food safety, and environmental health. The rapid detection and quantification of antimicrobial resistance are important for both infectious disease control and public health threat assessment. Technologies such as flow cytometry can provide clinicians with the early information, they need for appropriate antibiotic treatment. At the same time, cytometry platforms facilitate the measurement of antibiotic-resistant bacteria in environments impacted by human activities, enabling assessment of their impact on watersheds and soils. This review focuses on the latest applications of flow cytometry for the detection of pathogens and antibiotic-resistant bacteria in both clinical and environmental samples. Novel antimicrobial susceptibility testing frameworks embedding flow cytometry assays can contribute to the implementation of global antimicrobial resistance surveillance systems that are needed for science-based decisions and actions.

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Section: The Landscape Of Rapid Methods For Antibiotic Susceptibility...mentioning

confidence: 99%

Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Măruţescu

2023

Microorganisms

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“…The following phenotypic methods have as common objective the development of analytical tools rapidly detecting the effects of an antibiotic. Highlighted techniques are flow cytometry (FC), [49][50][51][52][53][54] dual molecular recognition, and mass spectrometry.…”

Section: Newer Approaches For the Phenotypic Astmentioning

confidence: 99%

Antibiotic Resistance Detection in Pseudomonas aeruginosa: Recent Strategies, Advances, and Challenges

Clarindo Lopes,

Koushanpour,

Wiebe

et al. 2024

Analysis & Sensing

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Antibiotic resistance presents a worldwide public health emergency, impeding the effective management of infectious diseases. Pseudomonas aeruginosa plays a substantial role as a bacterial pathogen, particularly in infections among hospitalized individuals, and those with weakened immune systems. Timely and accurate detection of antimicrobial resistance in P. aeruginosa is crucial for initiating tailored antibiotic therapy promptly, thus improving patient outcomes. Nevertheless, this endeavor encounters challenges due to the intricate and varied nature of antibiotic‐resistant strains. Extensive efforts have been invested in developing sensors and instrumentations for assessing antibiotic resistance or susceptibility (AST), aiming to enable personalized patient treatment with appropriate medications. This paper focuses on recent advancements in these methodologies, utilized for evaluating the degree of antibiotic resistance or susceptibility inpathogenic bacteria. Flow cytometry, dual molecular recognition and mass spectrometry are presented as newer phenotypic AST techniques. Genomic methods and the pyocyanin detection are listed as methodologies for the detection of resistance indicators.

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“…To speed up the detection, many studies have developed detection methods based on flow cytometry, DNA, Raman spectroscopy, polymerase chain reaction, etc. [ 4 , 5 , 6 , 7 ]. However, these methods also significantly increase the cost of analysis due to the expensive instruments.…”

Section: Introductionmentioning

confidence: 99%

YOLO Algorithm for Long-Term Tracking and Detection of Escherichia Coli at Different Depths of Microchannels Based on Microsphere Positioning Assistance

Sun

Rao

et al. 2022

Sensors

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The effect evaluation of the antibiotic susceptibility test based on bacterial solution is of great significance for clinical diagnosis and prevention of antibiotic abuse. Applying a microfluidic chip as the detection platform, the detection method of using microscopic images to observe bacteria under antibiotic can greatly speed up the detection time, which is more suitable for high-throughput detection. However, due to the influence of the depth of the microchannel, there are multiple layers of bacteria under the focal depth of the microscope, which greatly affects the counting and recognition accuracy and increases the difficulty of relocation of the target bacteria, as well as extracting the characteristics of bacterial liquid changes under the action of antibiotics. After the focal depth of the target bacteria is determined, although the z-axis can be controlled with the help of a three-dimensional micro-operator, the equipment is difficult to operate and the long-term changes of the target bacteria cannot be tracked quickly and accurately. In this paper, the YOLOv5 algorithm is adopted to accurately identify bacteria with different focusing states of multi-layer bacteria at the z-axis with any focal depth. In the meantime, a certain amount of microspheres were mixed into bacteria to assist in locating bacteria, which was convenient for tracking the growth state of bacteria over a long period, and the recognition rates of both bacteria and microspheres were high. The recognition accuracy and counting accuracy of bacteria are 0.734 and 0.714, and the two recognition rates of microspheres are 0.910 and 0.927, respectively, which are much higher than the counting accuracy of 0.142 for bacteria and 0.781 for microspheres with the method of enhanced depth of field (EDF method). Moreover, during long-term bacterial tracking and detection, target bacteria at multiple z-axis focal depth positions can be recorded by the aid of microspheres as a positioning aid for 3D reconstruction, and the focal depth positions can be repositioned within 3–10 h. The structural similarity (SSIM) of microscopic image structure differences at the same focal depth fluctuates between 0.960 and 0.975 at different times, and the root-mean-square error (RMSE) fluctuates between 8 and 12, which indicates that the method also has good relocation accuracy. Thus, this method provides the basis for rapid, high-throughput, and long-term analysis of microscopic changes (e.g., morphology, size) of bacteria detection under the addition of antibiotics with different concentrations based on microfluidic channels in the future.

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MICy: a Novel Flow Cytometric Method for Rapid Determination of Minimal Inhibitory Concentration

Cited by 7 publications

References 32 publications

Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Antibiotic Resistance Detection in Pseudomonas aeruginosa: Recent Strategies, Advances, and Challenges

YOLO Algorithm for Long-Term Tracking and Detection of Escherichia Coli at Different Depths of Microchannels Based on Microsphere Positioning Assistance

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