3× multiplexed detection of antibiotic resistant plasmids with single molecule sensitivity

Meena, Gopikrishnan G.; Hanson, Robert L.; Wood, Ryan L.; Brown, O.; Stott, Matthew A.; Robison, Richard A.; Pitt, William G.; Hawkins, Aaron R.; Schmidt, Holger

doi:10.1039/d0lc00640h

Cited by 17 publications

(9 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Specifically, a double stranded (ds) DNA sample corresponding to an antibiotic resistant pUC19- New Delhi metallo- β -lactamase (NDM) plasmid was prepared by sequence specifically extracting the target DNAs from the blood sample spiked with E. coli cells having pUC19-NDM plasmids and then labeled with nucleic acid staining dye. The target sample was prepared with a multi component system, consisting of cell extraction from the whole blood, nucleic acid sample preparation in a microfluidic chip [ 21 ]. While the initial plasmid concentration spiked into the starting amount of 7mL whole blood was known (5 ×10 6 CFU mL −1 ), there was non-trivial loss of DNA targets before the sample was introduced in the ARROW chip, in particular in the intermediate sample prep chip.…”

Section: Resultsmentioning

confidence: 99%

“…ARROW devices have shown efficient and amplification-free detection of single bioparticles while they flow through a microchannel under a mechanical or electrical driving force [ 7 ], [ 17 ], [ 18 ]. Moreover, multi-spot excitation for spectrally and spatially multiplexed target detection with excellent signal to noise (SNR) ratio was implemented with multimode interference (MMI) waveguides, [ 19 ]–[ 21 ]. Moreover, this platform can be easily integrated with a programmable bio sample preparation system for efficient and contamination-free sample transfer process to the sensing region [ 22 ], [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

FPGA Integrated Optofluidic Biosensor for Real-Time Single Biomarker Analysis

et al. 2022

View full text Add to dashboard Cite

Integrated optofluidic biosensors can fill the need for sensitive, amplification-free, multiplex single molecule detection which is relevant for containing the spread of infectious diseases such as COVID-19. Here, we demonstrate a rapid sample-to-answer scheme that uses a field programmable gate array (FPGA) to enable live monitoring of single particle fluorescence analysis on an optofluidic chip. Fluorescent nanobeads flowing through a micro channel are detected with 99% accuracy and particle concentrations in clinically relevant ranges from 3.4×10 4 to 3.4 × 10 6 /ml are determined within seconds to a few minutes without the need for post-experiment data extraction and analysis. In addition, other extract salient experimental parameters such as dynamic flow rate changes can be monitored in real time. The sensor is validated with real-time fluorescence detection of single bacterial plasmid DNA at attomolar concentrations, showing excellent promise for implementation as a point of care (POC) diagnostic tool.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

FPGA Integrated Optofluidic Biosensor for Real-Time Single Biomarker Analysis

et al. 2022

View full text Add to dashboard Cite

show abstract

“…Integrating sample processing and preconcentration on-chip can address these issues. Meena et al [ 44 ] used on-chip preconcentration followed by labeling and single-particle counting to quantify specific DNA sequences that encode antibiotic resistance in bacterial infections. They used one device to preconcentrate, isolate, and fluorescently label a DNA target from mixed samples using a monolith column with affixed capture probes.…”

Section: Spectroscopic Detectionmentioning

confidence: 99%

Advances in multiplex electrical and optical detection of biomarkers using microfluidic devices

Mitchell

Esene

2021

Anal Bioanal Chem

View full text Add to dashboard Cite

Graphical abstract Microfluidic devices can provide a versatile, cost-effective platform for disease diagnostics and risk assessment by quantifying biomarkers. In particular, simultaneous testing of several biomarkers can be powerful. Here, we critically review work from the previous 4 years up to February 2021 on developing microfluidic devices for multiplexed detection of biomarkers from samples. We focus on two principal approaches: electrical and optical detection methods that can distinguish and quantify biomarkers. Both electrical and spectroscopic multiplexed detection strategies are being employed to reach limits of detection below clinical sample levels. Some of the most promising strategies for point-of-care assays involve inexpensive materials such as paper-based microfluidic devices, or portable and accessible detectors such as smartphones. This review does not comprehensively cover all multiplexed microfluidic biomarker studies, but rather provides a critical evaluation of key work and suggests promising prospects for future advancement in this field. Electrical and optical multiplexing are powerful approaches for microfluidic biomarker analysis.

show abstract

“…The markers fluoresce in this light, and the fluorescence can be detected and measured by a photodetector as a signal for target presence. Targets detectable by this biosensor platform include antigens, cancer biomarkers, liposomes, nucleic acids, proteins, ribosomes, and virions [1][2][3][4][5][6].…”

Section: Introductionmentioning

confidence: 99%

Performance Comparison of Flow-Through Optofluidic Biosensor Designs

et al. 2021

View full text Add to dashboard Cite

Optofluidic flow-through biosensors are being developed for single particle detection, particularly as a tool for pathogen diagnosis. The sensitivity of the biosensor chip depends on design parameters, illumination format (side vs. top), and flow configuration (parabolic, two- and three-dimensional hydrodynamic focused (2DHF and 3DHF)). We study the signal differences between various combinations of these design aspects. Our model is validated against a sample of physical devices. We find that side-illumination with 3DHF produces the strongest and consistent signal, but parabolic flow devices process a sample volume more quickly. Practical matters of optical alignment are also discussed, which may affect design choice.

show abstract

3× multiplexed detection of antibiotic resistant plasmids with single molecule sensitivity

Abstract: Bacterial pathogens resistant to antibiotics have become a serious health threat. Those species which have developed resistance against multiple drugs such as the carbapenems, are more lethal as these are...

Cited by 17 publications

References 40 publications

FPGA Integrated Optofluidic Biosensor for Real-Time Single Biomarker Analysis

FPGA Integrated Optofluidic Biosensor for Real-Time Single Biomarker Analysis

Advances in multiplex electrical and optical detection of biomarkers using microfluidic devices

Performance Comparison of Flow-Through Optofluidic Biosensor Designs

Contact Info

Product

Resources

About