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

Needs, Sarah; Pivetal, Jérémy; Hayward, Jessica H.; Kidd, Stephen P.; Lam, HoYin; Diep, Tai The; Gill, Kiran Dip; Woodward, Martin J.; Reis, Nuno M.; Edwards, Alexander D.

doi:10.1039/d2sd00138a

Cited by 5 publications

(3 citation statements)

References 61 publications

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“…38 Recently, moving microcapillary antibiotic susceptibility testing (mcAST) was introduced, which uses microcapillaries to identify E. coli , and a smartphone for observation of the results. 39 The markers employed in the early detection of urinary tract infection are mainly urine xanthine oxidase and myeloperoxidase. A microbiological study was conducted using VITEK, but this required skilled personnel and an appropriate laboratory set-up.…”

Section: Resultsmentioning

confidence: 99%

Market-ready U-AST kit: simple, fast, cost-effective solution for concurrently detecting urinary tract infection and antibiotic resistance

Behere,

Haldar

2024

Anal. Methods

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Section: Resultsmentioning

confidence: 99%

Market-ready U-AST kit: simple, fast, cost-effective solution for concurrently detecting urinary tract infection and antibiotic resistance

Behere,

Haldar

2024

Anal. Methods

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“…In many systems, the detection of biomolecules occurs via the indirect detection of colour or fluorescence changes, with the most frequent examples including enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) but also including many other biochemical reactions or interactions. These have included the detection of specific bacteria and pathogens [53], the presence of proteins or nucleic acid biomarkers [38,40,52], and the monitoring of mammalian [37] and bacterial cell growth [59]. These assays and biosensors can be miniaturised using microfluidics and then monitored using open-source hardware systems.…”

Section: Colour and Fluorescence Detection In Microfluidicsmentioning

confidence: 99%

Open Hardware for Microfluidics: Exploiting Raspberry Pi Singleboard Computer and Camera Systems for Customisable Laboratory Instrumentation

Sarıyer,

Edwards,

Needs

2023

Biosensors

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The integration of Raspberry Pi miniature computer systems with microfluidics has revolutionised the development of low-cost and customizable analytical systems in life science laboratories. This review explores the applications of Raspberry Pi in microfluidics, with a focus on imaging, including microscopy and automated image capture. By leveraging the low cost, flexibility and accessibility of Raspberry Pi components, high-resolution imaging and analysis have been achieved in direct mammalian and bacterial cellular imaging and a plethora of image-based biochemical and molecular assays, from immunoassays, through microbial growth, to nucleic acid methods such as real-time-qPCR. The control of image capture permitted by Raspberry Pi hardware can also be combined with onboard image analysis. Open-source hardware offers an opportunity to develop complex laboratory instrumentation systems at a fraction of the cost of commercial equipment and, importantly, offers an opportunity for complete customisation to meet the users’ needs. However, these benefits come with a trade-off: challenges remain for those wishing to incorporate open-source hardware equipment in their own work, including requirements for construction and operator skill, the need for good documentation and the availability of rapid prototyping such as 3D printing plus other components. These advances in open-source hardware have the potential to improve the efficiency, accessibility, and cost-effectiveness of microfluidic-based experiments and applications.

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“…Thus, there is a pressing need for the development of novel AST methodologies that can detect all resistance mechanisms and provide rapid, precise and consistent results to better inform antibiotic treatment strategies [8,12]. There have been several attempts at developing such methods in the last decade with a variety of approaches [13][14][15][16][17][18][19][20][21][22]. These rapid phenotypic methods suffer from inherent limitations that prevent them from becoming more widely adopted, including cost, reliance on databases, narrow antibiotic ranges and high false positive rates [23][24][25][26].…”

Section: Introductionmentioning

confidence: 99%

Multipad agarose plate: a rapid and high-throughput approach for antibiotic susceptibility testing

Kals,

Mancini,

Kotar

et al. 2024

J. R. Soc. Interface.

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We describe a phenotypic antibiotic susceptibility testing (AST) method that can provide an eightfold speed-up in turnaround time compared with the current clinical standard by leveraging advances in microscopy and single-cell imaging. A newly developed growth plate containing 96 agarose pads, termed the multipad agarose plate (MAP), can be assembled at low cost. Pads can be prepared with dilution series of antibiotics. Bacteria are seeded on the pads and automatically imaged using brightfield microscopy, with a fully automated segmentation pipeline quantifying microcolony formation and growth rate. Using a test set of nine antibiotics with very different targets, we demonstrate that accurate minimum inhibitory concentration (MIC) measurements can be performed based on the growth rate of microcolonies within 3 h of incubation with the antibiotic when started from exponential phase. Faster, reliable and high-throughput methods for AST, such as MAP, could improve patient care by expediting treatment initiation and alleviating the burden of antimicrobial resistance.

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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

Abstract: Point-of-care antibiotic susceptibility test for urinary tract infections.

Cited by 5 publications

References 61 publications

Market-ready U-AST kit: simple, fast, cost-effective solution for concurrently detecting urinary tract infection and antibiotic resistance

Market-ready U-AST kit: simple, fast, cost-effective solution for concurrently detecting urinary tract infection and antibiotic resistance

Open Hardware for Microfluidics: Exploiting Raspberry Pi Singleboard Computer and Camera Systems for Customisable Laboratory Instrumentation

Multipad agarose plate: a rapid and high-throughput approach for antibiotic susceptibility testing

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