A Novel Microfluidic Assay for Rapid Phenotypic Antibiotic Susceptibility Testing of Bacteria Detected in Clinical Blood Cultures

Malmberg, Christer; Yuen, Pikkei; Spaak, Johanna; Cars, Otto; Tängdén, Thomas; Lagerbäck, Pernilla

doi:10.1371/journal.pone.0167356

Cited by 35 publications

(25 citation statements)

References 17 publications

(20 reference statements)

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“…overloading) without compromising fluid flow. Consistent with our previous study (17), there was no fluid flow through the gel-filled growth chamber. Instead, molecules travel through the growth chamber between the two flanking flow channels by diffusion.…”

Section: Design and Manufacture Of The Fluidic Systemsupporting

confidence: 92%

“…We have previously designed and evaluated a rapid phenotypic antibiotic susceptibility test where bacteria are mixed with agarose to form a gel within a growth chamber inside a microfluidic system, and subjected to a gradient of an antibiotic to enable MIC determination (17). The aim of the current study was to develop a high-throughput system (based on our previous design) capable of simultaneously analyzing 8 samples in one chip and with the possibility of testing several antibiotics in parallel.…”

Section: Introductionmentioning

confidence: 99%

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A high-throughput fluidic chip for rapid phenotypic antibiotic susceptibility testing

Wistrand-Yuen

Malmberg

Fatsis-Kavalopoulos

et al. 2019

Preprint

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Many patients with severe infections receive inappropriate empirical treatment and rapid detection of bacterial antibiotic susceptibility can in this context improve clinical outcome and reduce mortality. We have to this end developed a high-throughput fluidic chip for rapid phenotypic antibiotic susceptibility testing of bacteria. A total of 21 clinical isolates of Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus were acquired from the EUCAST Development Laboratory and tested against amikacin, ceftazidime and meropenem (Gramnegative bacteria) or gentamicin, ofloxacin and tetracycline (Gram-positive bacteria). The bacterial samples were mixed with agarose and loaded in 8 separate growth chambers in the fluidic chip. The chip was thereafter connected to a reservoir lid containing different antibiotics and a pump used to draw growth media with or without antibiotics into the chip for generation of diffusion-limited antibiotic gradients in the growth chambers. Bacterial microcolony growth was monitored using darkfield time-lapse microscopy and quantified using a cluster image analysis algorithm. Minimum inhibitory concentration (MIC) values were automatically obtained by tracking the growth rates of individual microcolonies in different regions of antibiotic gradients. Stable MIC values were obtained within 2-4 hours and the results showed categorical agreement to reference MIC values as determined with broth microdilution in 86% of the cases. ImportancePrompt and effective antimicrobial therapy is crucial for the management of patients with severe bacterial infections but is becoming increasingly difficult to provide due to emerging antibiotic resistance. The traditional methods for antibiotic susceptibility testing (AST) used in most clinical laboratories are reliable but slow with turnaround times of 2-3 days, which necessitates the use of empirical therapy with broad-spectrum antibiotics. There is a great need for fast and reliable AST methods that enable start of targeted treatment within a few hours to improve patient outcome and reduce overuse of broad-spectrum antibiotics. The high-throughput fluidic chip for phenotypic AST described in the present study enables data on antimicrobial resistance within 2-4 hours allowing for an early initiation of appropriate antibiotic therapy.

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Section: Design and Manufacture Of The Fluidic Systemsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

A high-throughput fluidic chip for rapid phenotypic antibiotic susceptibility testing

Wistrand-Yuen

Malmberg

Fatsis-Kavalopoulos

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…We have previously designed and evaluated a rapid phenotypic antibiotic susceptibility test where bacteria are mixed with agarose to form a gel within a growth chamber inside a microfluidic system and subjected to a gradient of an antibiotic to enable MIC determination (17). The aim of the current study was to develop a higher throughput system (based on our previous design) capable of simultaneously analyzing eight samples on one chip and with the possibility of testing several antibiotics in parallel.…”

mentioning

confidence: 99%

A Multiplex Fluidic Chip for Rapid Phenotypic Antibiotic Susceptibility Testing

Wistrand-Yuen

Malmberg

Fatsis-Kavalopoulos

et al. 2020

mBio

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Many patients with severe infections receive inappropriate empirical treatment, and rapid detection of bacterial antibiotic susceptibility can improve clinical outcome and reduce mortality. To this end, we have developed a multiplex fluidic chip for rapid phenotypic antibiotic susceptibility testing of bacteria. A total of 21 clinical isolates of Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus were acquired from the EUCAST Development Laboratory and tested against amikacin, ceftazidime, and meropenem (Gram-negative bacteria) or gentamicin, ofloxacin, and tetracycline (Gram-positive bacteria). The bacterial samples were mixed with agarose and loaded in an array of growth chambers in the chip where bacterial microcolony growth was monitored over time using automated image analysis. MIC values were automatically obtained by tracking the growth rates of individual microcolonies in different regions of antibiotic gradients. Stable MIC values were obtained within 2 to 4 h, and the results showed categorical agreement with reference MIC values as determined by broth microdilution in 86% of the cases. IMPORTANCE Prompt and effective antimicrobial therapy is crucial for the management of patients with severe bacterial infections but is becoming increasingly difficult to provide due to emerging antibiotic resistance. The traditional methods for antibiotic susceptibility testing (AST) used in most clinical laboratories are reliable but slow with turnaround times of 2 to 3 days, which necessitates the use of empirical therapy with broad-spectrum antibiotics. There is a great need for fast and reliable AST methods that enable starting targeted treatment within a few hours to improve patient outcome and reduce the overuse of broad-spectrum antibiotics. The multiplex fluidic chip for phenotypic AST described in the present study may enable data on antimicrobial resistance within 2 to 4 h, allowing for an early initiation of appropriate antibiotic therapy.

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“…The recently introduced technology by Accelerate Diagnostics uses morphokinetic cellular analysis to measure the capacity of a microbe to proliferate in the presence of an antibiotic [6]. At present, there is evidence that increasing the speed of the detection of antibiotic resistance is a real topic [7], [8,9], [10], [11]. In routine clinical microbiology settings, the time to measure antibiograms from an automated or manual system ranges from 6 to 12 hours.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of bacterial proliferation with a microfluidic-based device: Antibiochip

Pizzi¹,

Gallo²,

Ruiba³

et al. 2019

Preprint

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The measurement of the proliferation (and the relevant inhibition of proliferation) of microbes is used in different settings, from industry to laboratory medicine. Thus, in this study, the capacity of the Antibiochip (ELTEK spa), a microfluidic-based device, to measure the amount of E. coli in certain culture conditions, was evaluated. An Antibiochip is composed of V-shaped microchannels, and the amount of microparticles (such as microbes) is measured by the surface of the pellet after centrifugation. In the present study, different geometries, volumes and times were analyzed. When the best conditions were identified, serial dilutions of microbial cultures were tested to validate the linearity of the results. Then, with the use of wild E. coli strains isolated from medical samples, the relationship between bacterial susceptibility to antibiotics (gentamicin, amikacin and ceftriaxone) measured by standard methods and that measured by the Antibiochip was evaluated. In this report, the good quality performances of the methods, their linearity and the capacity to identify susceptible microbial strains after 60 minutes of incubation are shown. These results represent a novel approach for ultrarapid antibiograms in clinics.

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A Novel Microfluidic Assay for Rapid Phenotypic Antibiotic Susceptibility Testing of Bacteria Detected in Clinical Blood Cultures

Cited by 35 publications

References 17 publications

A high-throughput fluidic chip for rapid phenotypic antibiotic susceptibility testing

A high-throughput fluidic chip for rapid phenotypic antibiotic susceptibility testing

A Multiplex Fluidic Chip for Rapid Phenotypic Antibiotic Susceptibility Testing

Evaluation of bacterial proliferation with a microfluidic-based device: Antibiochip

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