Bacteria encapsulation and rapid antibiotic susceptibility test using a microfluidic microwell device integrating surface-enhanced Raman scattering

Huang, Hsiu-Kang; Cheng, Ho-Wen; Liao, Cheng-Chieh; Lin, Shang-Jyun; Chen, Yi‐Zih; Wang, Juen-Kai; Wang, Yuh‐Lin; Huang, Nien-Tsu

doi:10.1039/d0lc00425a

Cited by 32 publications

(24 citation statements)

References 30 publications

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“…In addition to the above methods, surface-enhanced Raman spectroscopy (SERS) is also widely applied for bacteria identification or ASTs. 12,13,28,29 Since the bacterial SERS signal originates from its released purines and their derivatives, 30,31 it possesses several advantages: (1) it is label-free, (2) it is non-invasive and (3) it reflects physiological activity. These advantages favor the integration of a SERS-based detection method with a multiplex microfluidic platform for high-throughput AST if the performance of the SERS-active substrate is uniform and reliable.…”

Section: Introductionmentioning

confidence: 99%

An antibiotic concentration gradient microfluidic device integrating surface-enhanced Raman spectroscopy for multiplex antimicrobial susceptibility testing

et al. 2022

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Section: Introductionmentioning

confidence: 99%

An antibiotic concentration gradient microfluidic device integrating surface-enhanced Raman spectroscopy for multiplex antimicrobial susceptibility testing

et al. 2022

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“…19 Alternatively, devices with integrated cultivation chambers or wells have been introduced. [20][21][22][23] Here, the chamber number is fixed, given by the initial design of the device. Isolation, i.e., closing the chamber, is achieved by valves 24,25 or again, by supply of non-water miscible fluids (oils).…”

Section: Introductionmentioning

confidence: 99%

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

Jusková

Schmitt

Kling

et al. 2021

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Rapid 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.

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“…As the pathogenic microbes turn noxious or resistant to drugs nowadays, the turnaround time to prescribe the correct antibiotics is a crucial matter to determine life or death. The current gold standard of antimicrobial susceptibility testing (AST) heavily relies on bacterial culture methods, such as broth microdilution [1] or disc diffusion [2], which require a long turnaround time (>24 h) [3,4] and a high initial bacterial count (at least 10 7 CFU/mL) [3]. Currently, the application of commercial instruments (e.g., Biomerieux Vitek2 compact and BD Phoenix™ Automated Microbiology System) in clinical laboratory routines can save time by replacing labor-intensive work with automation.…”

Section: Introductionmentioning

confidence: 99%

Developing Rapid Antimicrobial Susceptibility Testing for Motile/Non-Motile Bacteria Treated with Antibiotics Covering Five Bactericidal Mechanisms on the Basis of Bead-Based Optical Diffusometry

2020

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Rapid antimicrobial susceptibility testing (AST) is an effective measure in the treatment of infections and the prevention of bacterial drug resistance. However, diverse antibiotic types and bacterial characteristics have formed complicated barriers to rapid diagnosis. To counteract these limitations, we investigated the interactions between antibiotic-treated bacteria and functionalized microbeads in optical diffusometry. The conjugation with bacteria increased the effective microbead complex size, thereby resulting in a temporal diffusivity change. The yielded data were sorted and analyzed to delineate a pattern for the prediction of antimicrobial susceptibility. The outcome showed that a completed rapid AST based on the trend of microbead diffusivity could provide results within 3 h (2 h measurement + 1 h computation). In this research, we studied four bacterial strains, including Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus, and six antibiotics. Despite the different inhibitory effects caused by various antibiotics, similar trends in diffusivity alteration for all susceptible and resistant cases in the last 40 min of the 2-h measurement period were deduced. In addition, the AST results obtained using optical diffusometry showed good agreement with those acquired from the commercial instrument and conventional culture methods. Finally, we conducted a single-blinded clinical test, and the sensitivity, specificity, and accuracy of the system reached 92.9%, 91.4%, and 91.8%, respectively. Overall, the developed optical diffusometry showcased rapid AST with a small sample volume (20 μL) and low initial bacterial count (105 CFU/mL). This technique provided a promising way to achieve early therapy against microbial diseases in the future.

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Bacteria encapsulation and rapid antibiotic susceptibility test using a microfluidic microwell device integrating surface-enhanced Raman scattering

Abstract: We developed a microfluidic microwell device integrating SERS substrate for an efficient bacteria encapsulation and enrichment followed by in situ SERS-AST measurement, which can potentially apply for high throughput and multi-parallel AST.

Cited by 32 publications

References 30 publications

An antibiotic concentration gradient microfluidic device integrating surface-enhanced Raman spectroscopy for multiplex antimicrobial susceptibility testing

An antibiotic concentration gradient microfluidic device integrating surface-enhanced Raman spectroscopy for multiplex antimicrobial susceptibility testing

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

Developing Rapid Antimicrobial Susceptibility Testing for Motile/Non-Motile Bacteria Treated with Antibiotics Covering Five Bactericidal Mechanisms on the Basis of Bead-Based Optical Diffusometry

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