Combinatorial screening SlipChip for rapid phenotypic antimicrobial susceptibility testing

Li, Xiang; Liu, Xu; Yu, Ziqing; Luo, Yang; Hu, Qixin; Xu, Zhenye; Dai, Jia; Wu, Nannan; Shen, Feng

doi:10.1039/d2lc00661h

Cited by 11 publications

(9 citation statements)

References 49 publications

(55 reference statements)

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“…We have previously demonstrated a parallel multistep digital analysis (PAMDA) SlipChip device that can add additional reagents in parallel to the droplet to change the microenvironment . We have also demonstrated that a gradient can be formed or a reagent can be preloaded into the device for further screening. , Therefore, the SlipChip has potential as a tool for many digital biosensing applications.…”

Section: Resultsmentioning

confidence: 99%

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Rapid Bacteriophage Quantification by Digital Biosensing on a SlipChip Microfluidic Device

Li,

Hu,

Liu

et al. 2023

Anal. Chem.

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The rise of antimicrobial resistance (AMR) is a major global public health concern, and it is urgent to develop new antimicrobial drugs and alternative therapies. There has been growing interest in the use of phage therapy as an alternative to treat AMR, and it has shown promising results in early studies and clinical trials. Phage quantification is a crucial step in the development and application of phage therapy. The traditional double-layer plaque assay requires cumbersome manual operations and typically takes up to 18 h to yield a rough phage estimation. Spectrophotometry, flow cytometry, and PCR-based methods cannot distinguish between infectious and noninfectious phages. Here, we developed a digital biosensing method for rapid bacteriophage quantification on a digital phage SlipChip (dp-SlipChip) microfluidic device containing 2304 microdroplets in 3 nL. By compartmentalizing the phages and bacteria in nanoliter droplets and analyzing the growth profile of bacteria at 3 h, the number of infectious phages can be precisely quantified. The results from the dp-SlipChip were consistent with the traditional double-layer plaque assay method and exhibited higher consistency and repeatability. The dp-SlipChip does not require a complex fluidic handling instrument to generate and manipulate droplets. This SlipChip-based digital biosensing method not only provides a promising tool for rapid phage quantification, which is important for the use of phages in clinical practice to treat antimicrobial-resistant bacteria, but can also be used as an ultrasensitive, high-specificity method to detect bacteria. Furthermore, this approach can be applied to other digital biology studies that require analysis at the single-object level.

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Section: Resultsmentioning

confidence: 99%

“…54 We have also demonstrated that a gradient can be formed or a reagent can be preloaded into the device for further screening. 55,56 Therefore, the SlipChip has potential as a tool for many digital biosensing applications.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Rapid Bacteriophage Quantification by Digital Biosensing on a SlipChip Microfluidic Device

Li,

Hu,

Liu

et al. 2023

Anal. Chem.

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“…This is achieved by monitoring bacterial growth within nanoliter-sized droplets using bright-field imaging and analyzing alterations in bacterial quantity and morphology. 61 Zhao et al 62 introduced an innovative microfluidic platform that employs microchips with wettability patterns and programmed droplet manipulations to efficiently assemble diverse droplets. This platform facilitated the screening of cells in a high-throughput manner, allowing for the evaluation of different combinations of biochemical and mechanical cues.…”

Section: Immunoassaymentioning

confidence: 99%

“…Furthermore, the same group presented a combinatorial‐screening SlipChip that enables high‐throughput phenotypic AST within a 3‐h period. This is achieved by monitoring bacterial growth within nanoliter‐sized droplets using bright‐field imaging and analyzing alterations in bacterial quantity and morphology 61 …”

Section: Microfluidic Mechanisms For Pdmmentioning

confidence: 99%

Programmable droplet microfluidics for complex multistep bioassays

Huang,

Wang,

Cai

et al. 2024

Interdisciplinary Medicine

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Programmable droplet microfluidics (PDM) refers to a microfluidic device that integrates the functionality of a series of droplets with distinct spatial locations in a designated temporal order. PDM streamlines the intricate workflow of complex bioassays by enabling programmable and macroscopic droplet displacements, in which the droplets serve as reservoirs for reagents, microvalves for liquid insulation, and in some cases micropumps for mass transportation. As these droplets are intangible structures, the need for expensive microfabrication procedures is eliminated. Furthermore, the parallelization of the droplet series provides flexibility in controlling the throughput of the microfluidic analysis system. This paper provides a comprehensive review of PDMs enabled by various microfluidic mechanisms, including magnetism actuation, relative liquid displacement, capillary suction and sequential microdisplacement. Additionally, important applications of PDM systems for nucleic acid detection, immunoassay, drug testing, and sample recovery are also introduced. In conclusion, PDM demonstrates its potential as a highly advantageous tool for executing intricate multistep bioassays on a microfluidic platform. These technologies have exhibited superiority over their traditional counterparts in terms of size reduction, automation, and low sample/reagent consumption.

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“…3,4 Multiple studies have demonstrated the potential to miniaturise AST into microfluidic systems [5][6][7] and in single cell systems. [8][9][10] Single cell microscopy or individual particle tracking systems have reported the fastest phenotypic AST results, some in as little as 30 minutes [11][12][13] but most report results within 1-3 h. [14][15][16][17] Other methods to measure bacterial activity, including impedance, can also be used to categorise effects of antibiotics in 0.5-1 h. 18,19 However, there is often a trade-off between accuracy, accessibility and affordability. For example, rapid microscopy methods may require more expensive and bulkier optics compared to simpler growth readouts, and may be better suited to laboratory or centralised testing sites.…”

Section: Introductionmentioning

confidence: 99%

Moving microcapillary antibiotic susceptibility testing (mcAST) towards the clinic: unravelling kinetics of detection of uropathogenic E. coli, mass-manufacturing and usability for detection of urinary tract infections in human urine

et al. 2023

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Combinatorial screening SlipChip for rapid phenotypic antimicrobial susceptibility testing

Abstract: Antimicrobial resistance (AMR) by bacteria is a serious global threat, and a rapid, high-throughput, and easy-to-use phenotypic antimicrobial susceptibility testing (AST) method is essential for making timely treatment decisions and...

Cited by 11 publications

References 49 publications

Rapid Bacteriophage Quantification by Digital Biosensing on a SlipChip Microfluidic Device

Rapid Bacteriophage Quantification by Digital Biosensing on a SlipChip Microfluidic Device

Programmable droplet microfluidics for complex multistep bioassays

Moving microcapillary antibiotic susceptibility testing (mcAST) towards the clinic: unravelling kinetics of detection of uropathogenic E. coli, mass-manufacturing and usability for detection of urinary tract infections in human urine

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