Monitoring the growth and drug susceptibility of individual bacteria using asynchronous magnetic bead rotation sensors (original) (raw)
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Continuous growth of individual bacteria has been previously studied by direct observation using optical imaging. However, optical microscopy studies are inherently diffraction limited and limited in the number of individual cells that can be continuously monitored. Here we report on the use of the asynchronous magnetic bead rotation (AMBR) sensor, which is not diffraction limited. The AMBR sensor allows for the measurement of nanoscale growth dynamics of individual bacterial cells, over multiple generations. This torque-based magnetic bead sensor monitors variations in drag caused by the attachment and growth of a single bacterial cell. In this manner, we observed the growth and division of individual Escherichia coli, with 80-nm sensitivity to the cell length. Over the life cycle of a cell, we observed up to a 300% increase in the rotational period of the biosensor due to increased cell volume. In addition, we observed single bacterial cell growth response to antibiotics. This work demonstrates the non-microscopy limited AMBR biosensor for monitoring individual cell growth dynamics, including cell elongation, generation time, lag time, and division, as well as their sensitivity to antibiotics.
2012
The long turnaround time in antimicrobial susceptibility testing (AST) endangers patients and encourages the administration of wide spectrum antibiotics, thus resulting in alarming increases of multi-drug resistant pathogens. A method for faster detection of bacterial proliferation presents one avenue towards addressing this global concern. We report on a label-free asynchronous magnetic bead rotation (AMBR) based viscometry method that rapidly detects bacterial growth and determines drug sensitivity by measuring changes in the suspension's viscosity. With this platform, we observed the growth of a uropathogenic Escherichia coli isolate, with an initial concentration of 50 cells per drop, within 20 minutes; in addition, we determined the gentamicin minimum inhibitory concentration (MIC) of the E. coli isolate within 100 minutes. We thus demonstrated a label-free, micro-viscometer platform that can measure bacterial growth and drug susceptibility more rapidly, with lower initial bacterial counts than existing commercial systems, and potentially with any microbial strains.
Lab on a Chip, 2011
Inappropriate antibiotic use is a major factor contributing to the emergence and spread of antimicrobial resistance. The long turnaround time (over 24 hours) required for clinical antimicrobial susceptibility testing (AST) often results in patients being prescribed empiric therapies, which may be inadequate, inappropriate, or overly broad-spectrum. A reduction in the AST time may enable more appropriate therapies to be prescribed earlier. Here we report on a new diagnostic asynchronous magnetic bead rotation (AMBR) biosensor droplet microfluidic platform that enables single cell and small cell population growth measurements for applications aimed at rapid AST. We demonstrate the ability to rapidly measure bacterial growth, susceptibility, and the minimum inhibitory concentration (MIC) of a small uropathogenic Escherichia coli population that was confined in microfluidic droplets and exposed to concentrations above and below the MIC of gentamicin. Growth was observed below the MIC, and no growth was observed above the MIC. A 52% change in the sensor signal (i.e. rotational period) was observed within 15 minutes, thus allowing AST measurements to be performed potentially within minutes.
Analytical Chemistry, 2012
The long turnaround time in antimicrobial susceptibility testing (AST) endangers patients and encourages the administration of wide spectrum antibiotics, thus resulting in alarming increases of multidrug resistant pathogens. A method for faster detection of bacterial proliferation presents one avenue toward addressing this global concern. We report on a label-free asynchronous magnetic bead rotation (AMBR) based viscometry method that rapidly detects bacterial growth and determines drug sensitivity by measuring changes in the suspension's viscosity. With this platform, we observed the growth of a uropathogenic Escherichia coli isolate, with an initial concentration of 50 cells per drop, within 20 min; in addition, we determined the gentamicin minimum inhibitory concentration (MIC) of the E. coli isolate within 100 min. We thus demonstrated a labelfree, microviscometer platform that can measure bacterial growth and drug susceptibility more rapidly, with lower initial bacterial counts than existing commercial systems, and potentially with any microbial strains.
Self-Assembled Biosensors on a Solid Interface for Rapid Detection and Growth Monitoring of Bacteria
2012
Developing rapid methods for pathogen detection and growth monitoring at low cell and analyte concentrations is an important goal, which numerous technologies are working towards solving. Rapid biosensors have already made a dramatic impact on improving patient outcomes and with continued development, these technologies may also help limit the emergence of antimicrobial resistance and reduce the ever expanding risk of foodborne illnesses. One technology that is being developed with these goals in mind is asynchronous magnetic bead rotation (AMBR) biosensors. Self-assembled AMBR biosensors have been demonstrated at water/air and water/oil interfaces, and here, for the first time, we report on self-assembled AMBR biosensors used at a solid interface. The solid interface configuration was used to measure the growth of Escherichia coli with two distinct phenomena at low cell concentrations: firstly, the AMBR rotational period decreased and secondly, the rotational period increased after several division times. Taking advantage of this low cell concentration behavior, a 20 % signal change from the growth of E. coli O157:H7 was detected in 91 ± 4 minutes, with a starting concentration of 5 x 10 3 CFU/mL. Such a rapid cell growth sensor could dramatically improve the detection time and sensitivity in applications requiring phenotypic testing of target cells.
Rapid bacterial growth and antimicrobial response using self-assembled magnetic bead sensors
Sensors and Actuators B: Chemical, 2014
Despite advances in molecular diagnostics, phenotypic results remain essential, especially due to the ever increasing presence of antimicrobial resistance in healthcare-acquired infections and in foodborne illnesses. Since phenotypic information is still crucial, the objective of this work was to combine the robustness and cost-effectiveness of traditional bacterial culture, and to approach the speed of molecular diagnostic methods. With concentrations as low as 5 × 10 3 CFU/mL, the growth of pathogenic Escherichia coli O157:H7 was detected in 91 ± 4 min, which is on par with reported real-time PCR times used for food microbiology testing. Results were obtained using a 48 sensor prototype device that accepted standard well plates. A sensor with the reported rapid growth monitoring capabilities could dramatically improve the detection time and sensitivity in applications that require rapid phenotypic information, such as with patient samples and food testing.
2007
We report on a new technique which was used to detect single Escherichia coli that is based on the changes in the nonlinear rotation of a magnetic microsphere driven by an external magnetic field. The presence of one Escherichia Coli bacterium on the surface of a 2.0 µm magnetic microsphere caused an easily measurable change in the drag of the system and, therefore, in the nonlinear rotation rate. The straight-forward measurement uses standard microscopy techniques and the observed average shift in the nonlinear rotation rate changed by a factor of ∼3.8.
Micro-magnet arrays for specific single bacterial cell positioning
Journal of Magnetism and Magnetic Materials, 2015
In various contexts such as pathogen detection or analysis of microbial diversity where cellular heterogeneity must be taken into account, there is a growing need for tools and methods that enable microbiologists to analyze bacterial cells individually. One of the main challenges in the development of new platforms for single cell studies is to perform precise cell positioning, but the ability to specifically target cells is also important in many applications. In this work, we report the development of new strategies to selectively trap single bacterial cells upon large arrays, based on the use of micro-magnets. Escherichia coli bacteria were used to demonstrate magnetically driven bacterial cell organization. In order to provide a flexible approach adaptable to several applications in the field of microbiology, cells were magnetically and specifically labeled using two different strategies, namely immunomagnetic labeling and magnetic in situ hybridization. Results show that centimeter-sized arrays of targeted, isolated bacteria can be successfully created upon the surface of a flat magnetically patterned hard magnetic film. Efforts are now being directed towards the integration of a detection tool to provide a complete micro-system device for a variety of microbiological applications.
High frequency asynchronous magnetic bead rotation for improved biosensors
2010
Biosensors with increasingly high sensitivity are crucial for probing small scale properties. The asynchronous magnetic bead rotation ͑AMBR͒ sensor is an emerging sensor platform, based on magnetically actuated rotation. Here the frequency dependence of the AMBR sensor's sensitivity is investigated. An asynchronous rotation frequency of 145 Hz is achieved. This increased frequency will allow for a calculated detection limit of as little as a 59 nm change in bead diameter, which is a dramatic improvement over previous AMBR sensors and further enables physical and biomedical applications.