scholarly journals Real-time respiration changes as a viability indicator for rapid antibiotic susceptibility testing in a microfluidic chamber array

2021 ◽  
Author(s):  
Petra Jusková ◽  
Steven Schmitt ◽  
André Kling ◽  
Darius G. Rackus ◽  
Martin Held ◽  
...  
Keyword(s):  
Antibiotic Susceptibility ◽  
Bacterial Infections ◽  
Susceptibility Testing ◽  
Pathogen Resistance ◽  
Rapid Identification ◽  
Individual Growth ◽  
Diagnostic Process ◽  
Antibiotic Susceptibility Testing ◽  
Microfluidic Chamber ◽  
Antibiotic Compounds

ABSTRACTRapid identification of a pathogen and the measurement of its antibiotic susceptibility are key elements in the diagnostic process of bacterial infections. Microfluidic technologies offer great control over handling and manipulation of low sample volumes with the possibility to study microbial cultures on the single-cell level. Downscaling the dimensions of cultivation systems directly results in a lower number of bacteria required for antibiotic susceptibility testing (AST) and thus in a reduction of the time to result. The developed platform presented in this work allows the reading of pathogen resistance profiles within 2-3 hours based on the changes of the dissolved oxygen levels during bacterial cultivation. The platform contains hundreds of individual growth chambers prefilled with a hydrogel containing oxygen-sensing nanoprobes and different concentrations of antibiotic compounds. The performance of the microfluidic platform is tested using quality control Escherichia coli strains (ATCC 25922 and ATCC 35218) in response to different clinically relevant antibiotics. The achieved results are in agreement with values given in clinical reference guides and independent measurements using a clinical AST protocol. Finally, the platform is successfully used for AST of an E. coli clinical isolate obtained from a patient blood culture.

scholarly journals A Multiplex Fluidic Chip for Rapid Phenotypic Antibiotic Susceptibility Testing

mBio ◽  
2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Pikkei Wistrand-Yuen ◽  
Christer Malmberg ◽  
Nikos Fatsis-Kavalopoulos ◽  
Moritz Lübke ◽  
Thomas Tängdén ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
Antibiotic Susceptibility ◽  
Bacterial Infections ◽  
Susceptibility Testing ◽  
Empirical Therapy ◽  
Automated Image Analysis ◽  
Antibiotic Susceptibility Testing ◽  
Content Type ◽  
Broad Spectrum Antibiotics ◽  
Severe Infections

ABSTRACT 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.

scholarly journals A portable and high-integrated 3D microfluidic chip for bacterial quantification and antibiotic susceptibility testing

2021 ◽  
Author(s):  
Wenshuai Wu ◽  
Gaozhe Cai ◽  
Yang Liu
Keyword(s):  
Single Cell ◽  
Antibiotic Susceptibility ◽  
Bacterial Infections ◽  
Susceptibility Testing ◽  
Dynamic Range ◽  
Antibiotic Susceptibility Testing ◽  
Technical Potential ◽  
Uniform Sample ◽  
Pdms Chip ◽  
User Friendly

On-site single-cell antibiotic susceptibility testing (sc-AST) provides unprecedented technical potential to improve the treatment of bacterial infections and study heterogeneous resistance to antibiotics. Herein, we developed a portable and high-integrated 3D polydimethylsiloxane (PDMS) chip to perform fast and on-site bacteria quantification and sc-AST. The 3D arrangement of the chambers significantly improved the integration of reaction units (~10000/cm2) and widened the dynamic range to 5 orders of magnitude. A capillary valve-based flow distributor was adopted for flow equidistribution in 64 parallel channels and uniform sample loading in as short as 2 s. The degassed PDMS enabled this device to independently dispense the sample into 3D chamber array with almost 100% efficiency. The quantification of Escherichia coli (E. coli) strains with various activity was accomplished in 0.5-2 h, shortened by 20 h in comparison to the traditional plate counting. The functionality of our platform was demonstrated with several effective antibiotics by determining minimum inhibitory concentrations at single-cell level. Furthermore, we utilized the lyophilization of test reagents and needle-mediated reagents rehydration to realize one-step on-site sc-AST. The results indicate that the proposed sc-AST platform is portable, highly sensitive, fast, accurate and user-friendly, thus it has the potential to facilitate precise therapy in time and monitor the treatment. Meanwhile, it could serve as an approach for investigating the mechanisms of heteroresistance at single-cell resolution.

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

2019 ◽  
Author(s):  
Pikkei Wistrand-Yuen ◽  
Christer Malmberg ◽  
Nikos Fatsis-Kavalopoulos ◽  
Moritz Lübke ◽  
Thomas Tängdén ◽  
...  
Keyword(s):  
High Throughput ◽  
Broad Spectrum ◽  
Antibiotic Susceptibility ◽  
Bacterial Infections ◽  
Susceptibility Testing ◽  
Empirical Therapy ◽  
Growth Media ◽  
Antibiotic Susceptibility Testing ◽  
Broad Spectrum Antibiotics ◽  
Growth Chambers

AbstractMany 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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