Yating Chai

Rapid and sensitive detection of Salmonella Typhimurium on eggshells by using wireless biosensors.

This article presents rapid, sensitive, direct detection of Salmonella Typhimurium on eggshells by using wireless magnetoelastic (ME) biosensors. The biosensor consists of a freestanding, strip-shaped ME resonator as the signal transducer and the E2 phage as the biomolecular recognition element that selectively binds with Salmonella Typhimurium. This ME biosensor is a type of mass-sensitive biosensor that can be wirelessly actuated into mechanical resonance by an externally applied timevarying magnetic field. When the biosensor binds with Salmonella Typhimurium, the mass of the sensor increases, resulting in a decrease in the sensors resonant frequency. Multiple E2 phage-coated biosensors (measurement sensors) were placed on eggshells spiked with Salmonella Typhimurium of various concentrations (1.6 to 1.6 × 10(7) CFU/cm(2)). Control sensors without phage were also used to compensate for environmental effects and nonspecific binding. After 20 min in a humidity-controlled chamber (95%) to allow binding of the bacteria to the sensors to occur, the resonant frequency of the sensors was wirelessly measured and compared with their initial resonant frequency. The resonant frequency change of the measurement sensors was found to be statistically different from that of the control sensors down to 1.6 × 10(2) CFU/cm(2), the detection limit for this work. In addition, scanning electron microscopy imaging verified that the measured resonant frequency changes were directly related to the number of bound cells on the sensor surface. The total assay time of the presented methodology was approximately 30 min, facilitating rapid detection of Salmonella Typhimurium without any preceding sampling procedures.

Effects of surface functionalization on the surface phage coverage and the subsequent performance of phage-immobilized magnetoelastic biosensors

One of the important applications for which phage-immobilized magnetoelastic (ME) biosensors are being developed is the wireless, on-site detection of pathogenic bacteria for food safety and bio-security. Until now, such biosensors have been constructed by immobilizing a landscape phage probe on gold-coated ME resonators via physical adsorption. Although the physical adsorption method is simple, the immobilization stability and surface coverage of phage probes on differently functionalized sensor surfaces need to be evaluated as a potential way to enhance the detection capabilities of the biosensors. As a model study, a filamentous fd-tet phage that specifically binds streptavidin was adsorbed on either bare or surface-functionalized gold-coated ME resonators. The surface functionalization was performed through the formation of three self-assembled monolayers with a different terminator, based on the sulfur-gold chemistry: AC (activated carboxy-terminated), ALD (aldehyde-terminated), and MT (methyl-terminated). The results, obtained by atomic force microscopy, showed that surface functionalization has a large effect on the surface phage coverage (46.8%, 49.4%, 4.2%, and 5.2% for bare, AC-, ALD-, and MT-functionalized resonators, respectively). In addition, a direct correlation of the observed surface phage coverage with the quantity of subsequently captured streptavidin-coated microbeads was found by scanning electron microscopy and by resonance frequency measurements of the biosensors. The differences in surface phage coverage on the differently functionalized surfaces may then be used to pattern the phage probe layer onto desired parts of the sensor surface to enhance the detection capabilities of ME biosensors.

A surface-scanning coil detector for real-time, in-situ detection of bacteria on fresh food surfaces

Proof-in-principle of a new surface-scanning coil detector has been demonstrated. This new coil detector excites and measures the resonant frequency of free-standing magnetoelastic (ME) biosensors that may now be placed outside the coil boundaries. With this coil design, the biosensors are no longer required to be placed inside the coil before frequency measurement. Hence, this new coil enables bacterial pathogens to be detected on fresh food surfaces in real-time and in-situ. The new coil measurement technique was demonstrated using an E2 phage-coated ME biosensor to detect Salmonella typhimurium on tomato surfaces. Real-time, in-situ detection was achieved with a limit of detection (LOD) statistically determined to be lower than 1.5×10(3) CFU/mm(2) with a confidence level of difference higher than 95% (p<0.05).

Design of a surface-scanning coil detector for direct bacteria detection on food surfaces using a magnetoelastic biosensor

The real-time, in-situ bacteria detection on food surfaces was achieved by using a magnetoelastic biosensor combined with a surface-scanning coil detector. This paper focuses on the coil design for signal optimization. The coil was used to excite the sensors vibration and detect its resonant frequency signal. The vibrating sensor creates a magnetic flux change around the coil, which then produces a mutual inductance. In order to enhance the signal amplitude, a theory of the sensors mutual inductance with the measurement coil is proposed. Both theoretical calculations and experimental data showed that the working length of the coil has a significant effect on the signal amplitude. For a 1 mm-long sensor, a coil with a working length of 1.3 mm showed the best signal amplitude. The real-time detection of Salmonella bacteria on a fresh food surface was demonstrated using this new technology.

Surface-scanning coil detectors for magnetoelastic biosensors: A comparison of planar-spiral and solenoid coils

This research introduces a planar spiral coil as a surface-scanning detector for magnetoelastic biosensors, which have been used to detect bacteria directly on food surfaces. The planar coil was compared with the previously investigated solenoid coil, in terms of the magnetic flux change, signal amplitude, and detection distance. Both theoretical calculations and experimental results demonstrated that the planar coil detector yields a dramatically improved signal amplitude and greater detection distance. In addition, simultaneous measurement of multiple biosensors on surfaces was demonstrated. This planar coil is therefore anticipated to facilitate the detection of bacteria on surfaces using magnetoelastic biosensors.

Blocking Non-Specific Binding for Phage-Based Magnetoelastic Biosensors

The magnetoelastic (ME) biosensors are used to detect pathogen in fresh juice or milk by solenoid coil, and also developed for real-time, direct pathogen detection on food surfaces by surface-scanning coil. This paper presents blocking effect of different reagents on non-specific binding for detecting Salmonella typhimurium in apple juice using phage-based magnetoelastic biosensors. Three different blocking reagents of Bovine serum albumin, Superblock blocking buffer and blocker BLOTTO were used and evaluated. The results shows that blocker BLOTTO has the best blocking effect on non-specific binding.

Rapid, enhanced detection of Salmonella Typhimurium on fresh spinach leaves using micron-scale, phage-coated magnetoelastic biosensors

In order to cost-effectively and rapidly detect bacterial food contamination in the field, the potential usefulness of phage-coated magnetoelastic (ME) biosensors has been recently reported. These biosensors are freestanding, mass-sensitive biosensors that can be easily batch-fabricated, thereby reducing the fabrication cost per sensor to a fraction of a cent. In addition, the biosensors can be directly placed on fresh produce surfaces and used to rapidly monitor possible bacterial food contamination without any preceding sample preparation. Previous investigations showed that the limit of detection (LOD) with millimeter-scale ME biosensors was fairly low for fresh produce with smooth surfaces (e.g., tomatoes and shell eggs). However, the LOD is anticipated to be dependent on the size of the biosensors as well as the topography of produce surfaces of interest. This paper presents an investigation into the use of micron-scale, phage-coated ME biosensors for the enhanced detection of Salmonella Typhimurium on fresh spinach leaves.

In-situ detection of multiple pathogenic bacteria on food surfaces

Real-time in-situ detection of pathogenic bacteria on fresh food surfaces was accomplished with phage-based magnetoelastic (ME) biosensors. The ME biosensor is constructed of a small rectangular strip of ME material that is coated with a biomolecular recognition element (phage, antibodies or proteins, etc.) that is specific to the target pathogen. This mass-sensitive ME biosensor is wirelessly actuated into mechanical resonance by an externally applied time-varying magnetic field. When the biosensor binds with target bacteria, the mass of the sensor increases, resulting in a decrease in the sensors resonant frequency. In order to compensate for nonspecific binding, control biosensors without phage were used in this experiment. In previous research, the biosensors were measured one by one. However, the simultaneous measurement of multiple sensors was accomplished in this research, and promises to greatly shorten the analysis time for bacterial detection. Additionally, the use of multiple biosensors enables the possibility of simultaneous detection of different pathogenic bacteria. This paper presents results of experiments in which multiple phage-based ME biosensors were simultaneously monitored. The E2 phage and JRB7 phage from a landscape phage library served as the bio-recognition element that have the capability of binding specifically with Salmonella typhimurium and B. anthracis spores, respectively. Real-time in-situ detection of Salmonella typhimurium and B. anthracis spores on food surfaces are presented.

High throughput pathogen screening for food safety using magnetoelastic biosensors

In order to secure food safety, high throughput pathogen screening technique that can quickly identify and isolate unsafe contaminated foods has been long-desired. Recently, magnetoelastic (ME) free-standing biosensors have been investigated as a label-free wireless biosensor system for real-time pathogen detection. ME biosensor is composed of a ME resonator coated with a bio-molecular recognition element that binds specifically with a target pathogen. Once the biosensor comes into contact with the target pathogen, binding occurs, resulting in a decrease of the sensors resonant frequency. Interrogated through magnetic signals, large amount of ME sensors can be deployed and monitored wirelessly. ME biosensors have been investigated to detect foodborne pathogens in cultures and liquid foods. Recently, it has been demonstrated that phage-based ME biosensors are able to directly detect Salmonella Typhimurium on food surfaces without the requirement of pre-analysis culture preparation. This paper will review the novel ME biosensor technique, including the detection principle, the characterization of the sensor performance, the deployment of multiple sensor detection and their applications, especially for food safety analysis. The ME biosensor technique has the potential to be a powerful pathogen screening tool for detecting contaminated food, identifying critical hazard points, and tracking contamination sources along the entire food supply chain.

Self-propelled, phage-based magnetoelastic biosentinels for detection of pathogens in liquid

This paper presents the concept of self-propelled magnetoelastic (ME) biosentinels that seek out and capture pathogenic bacteria in stagnant liquids. These biosentinels are composed of a free-standing, asymmetric-shaped ME resonator coated with a filamentous landscape phage that specifically binds with a pathogen of interest. When a time-varying magnetic pulse is applied, the ME biosentinels can be placed into mechanical resonance by magnetostriction. The resultant asymmetric vibration then generates a net force on the surroundings and hence generates autonomous motion in the liquid. As soon as the biosentinels find and bind with the target pathogen through the phage-based biomolecular recognition, a change in the biosentinel’s resonant frequency occurs, and thereby the presence of the target pathogen can be detected. In order to actuate the ME biosentinels into mechanical resonance of a desired mode, modal analysis using the three-dimensional finite element method was performed. In addition, the design of a magnetic chamber that can control the orientation and/or translation of a biosentinel is discussed.