Evangelyn C. Alocilja

Market analysis of biosensors for food safety

This paper is presented as an overview of the pathogen detection industry. The review includes pathogen detection markets and their prospects for the future. Potential markets include the medical, military, food, and environmental industries. Those industries combined have a market size of

Design and fabrication of a microimpedance biosensor for bacterial detection

563 million for pathogen detecting biosensors and are expected to grow at a compounded annual growth rate of 4.5%. The food market is further segmented into different food product industries. The overall food-pathogen testing market is expected to grow to

Aptasensors for detection of microbial and viral pathogens

192 million and 34 million tests by 2005. The trend in pathogen testing emphasizes the need to commercialize biosensors for the food safety industry as legislation creates new standards for microbial monitoring. With quicker detection time and reusable features, biosensors will be important to those interested in real time diagnostics of disease causing pathogens. As the world becomes more concerned with safe food and water supply, the demand for rapid detecting biosensors will only increase.

A conductometric biosensor for biosecurity

A biosensor for bacterial detection was developed based on microelectromechanical systems, heterobifunctional crosslinkers and immobilized antibodies. The sensor detected the change in impedance caused by the presence of bacteria immobilized on interdigitated gold electrodes and was fabricated from (100) silicon with a 2-/spl mu/m layer of thermal oxide as an insulating layer. The sensor active area is 9.6 mm/sup 2/ and consists of two interdigital gold electrode arrays measuring 0.8 /spl times/ 6 mm. Escherichia coli specific antibodies were immobilized to the oxide between the electrodes to create a biological sensing surface. The impedance across the interdigital electrodes was measured after immersing the biosensor in solution. Bacteria cells present in the sample solution attached to the antibodies and became tethered to the electrode array, thereby causing a change in measured impedance. The biosensor was able to discriminate between different cellular concentrations from 10/sup 5/ to 10/sup 7/ CFU/mL in pure culture. The sample testing process, including data acquisition, required 5 min. The design, fabrication, and testing of the biosensor is discussed along with the implications of these findings toward further biosensor development.

Single-walled carbon nanotubes dispersed in aqueous media via non-covalent functionalization: Effect of dispersant on the stability, cytotoxicity, and epigenetic toxicity of nanotube suspensions

Abstract Aptamers are specific nucleic acid sequences that can bind to a wide range of non-nucleic acid targets with high affinity and specificity. These molecules are identified and selected through an in vitro process called SELEX (systematic evolution of ligands by exponential enrichment). Proteins are the most common targets in aptamer selection. In diagnostic and detection assays, aptamers represent an alternative to antibodies as recognition agents. Cellular detection is a promising area in aptamer research. One of its principal advantages is the ability to target and specifically differentiate microbial strains without having previous knowledge of the membrane molecules or structural changes present in that particular microorganism. The present review focuses on aptamers, SELEX procedures, and aptamer-based biosensors (aptasensors) for the detection of pathogenic microorganisms and viruses. Special emphasis is placed on nanoparticle-based platforms.

A Multichannel Femtoampere-Sensitivity Potentiostat Array for Biosensing Applications

The paper describes the development of a conductometric biosensor for detecting foodborne pathogens. The biosensor consists of two components: an immunosensor that is based on electrochemical sandwich immunoassay, and a reader for signal measurement. The architecture of the immunosensor utilizes a lateral flow system that allows the liquid sample to move from one pad to another. The biosensor provides a specific, sensitive, low volume, and near real-time detection mechanism. Results are presented to highlight the performance of the biosensor for enterohemorrhagic Escherichia coli O157:H7 and Salmonella spp., which are of concern to biosecurity. The lower limit of detection is approximately 7.9 x 10(1) colony forming units per milliliter within a 10-min process. The ability to change the specificity of the antibodies will enable the biosensor to be used as a detection device for other types of foodborne pathogens.

A microfabricated biosensor for detecting foodborne bioterrorism agents

As the range of applications for carbon nanotubes (CNTs) rapidly expands, understanding the effect of CNTs on prokaryotic and eukaryotic cell systems has become an important research priority, especially in light of recent reports of the facile dispersion of CNTs in a variety of aqueous systems including natural water. In this study, single-walled carbon nanotubes (SWCNTs) were dispersed in water using a range of natural (gum arabic, amylose, Suwannee River natural organic matter) and synthetic (polyvinyl pyrrolidone, Triton X-100) dispersing agents (dispersants) that attach to the CNT surface non-covalently via different physiosorption mechanisms. The charge and the average effective hydrodynamic diameter of suspended SWCNTs as well as the concentration of exfoliated SWCNTs in the dispersion were found to remain relatively stable over a period of 4 weeks. The cytotoxicity of suspended SWCNTs was assessed as a function of dispersant type and exposure time (up to 48 h) using general viability bioassay with Escherichia coli and using neutral red dye uptake (NDU) bioassay with WB-F344 rat liver epithelia cells. In the E. coli viability bioassays, three types of growth media with different organic loadings and salt contents were evaluated. When the dispersant itself was non-toxic, no losses of E. coli and WB-F344 viability were observed. The cell viability was affected only by SWCNTs dispersed using Triton X-100, which was cytotoxic in SWCNT-free (control) solution. The epigenetic toxicity of dispersed CNTs was evaluated using gap junction intercellular communication (GJIC) bioassay applied to WB-F344 rat liver epithelial cells. With all SWCNT suspensions except those where SWCNTs were dispersed using Triton X-100 (wherein GJIC could not be measured because the sample was cytotoxic), no inhibition of GJIC in the presence of SWCNTs was observed. These results suggest a strong dependence of the toxicity of SWCNT suspensions on the toxicity of the dispersant and point to the potential of non-covalent functionalization with non-toxic dispersants as a method for the preparation of stable aqueous suspensions of biocompatible CNTs.

Fluorescent bio-barcode DNA assay for the detection of Salmonella enterica serovar Enteritidis

Rapid and accurate detection of pathogens using conductometric biosensors requires potentiostats that can measure small variations in conductance. In this paper, we present an architecture and implementation of a multichannel potentiostat array based on a novel semi-synchronous sigma-delta (SigmaDelta) analog-to-digital conversion algorithm. The algorithm combines continuous time SigmaDelta with time-encoding machines, and enables measurement of currents down to femtoampere range. A 3-mmtimes3-mm chip implementing a 42-channel potentiostat array has been prototyped in a 0.5-mum CMOS technology. Measured results demonstrate that the prototype can achieve 10 bits of resolution, with a sensitivity down to 50-fA current. The power consumption of the potentiostat has been measured to be 11 muW per channel for a sampling rate of 250 kHz. Experiments with a conductometric biosensor specific to Bacillus Cereus bacterium, demonstrate the ability of the potentiostat in identifying different concentration levels of the pathogen in a biological sample

A multiplex nanoparticle-based bio-barcoded DNA sensor for the simultaneous detection of multiple pathogens ☆

A biosensor for the detection of pathogenic bacteria was developed for biosecurity applications. The sensor was fabricated using photolithography and incorporates heterobifunctional crosslinkers and immobilized antibodies. The sensor detected the change in impedance caused by the presence of bacteria immobilized on interdigitated gold electrodes and was fabricated from (100) silicon with a 2-/spl mu/m layer of thermal oxide as an insulating layer. The sensor has a large active area of 9.6 mm/sup 2/ and consists of two interdigital gold electrode arrays each measuring 0.8 /spl times/ 6 mm. Pathogenic Escherichia coli and Salmonella infantis were tested in serially diluted pure culture. Analyte specific antibodies were immobilized to the oxide between the electrodes to create a biological sensing surface. After immersing the biosensor in solution, the impedance across the interdigital electrodes was measured. Bacteria cells present in the sample solution attached to the antibodies and became tethered to the electrode array thereby causing a change in measured impedance. The biosensor was able to discriminate between different cellular concentrations from 10/sup 4/ - 10/sup 7/ CFU/mL (colony-forming units per milliliter) in solution. The sample testing process, including data acquisition, required 5 min. The design, fabrication, and testing of the biosensor is discussed along with the implications of these findings toward further biosensor development.

Electrically active polyaniline coated magnetic (EAPM) nanoparticle as novel transducer in biosensor for detection of Bacillus anthracis spores in food samples.

Salmonella enterica serovar Enteritidis is one of the most frequently reported causes of foodborne illness. It is a major threat to the food safety chain and public health. A highly amplified bio-barcode DNA assay for the rapid detection of the insertion element (Iel) gene of Salmonella Enteritidis is reported in this paper. The biosensor transducer is composed of two nanoparticles: gold nanoparticles (Au-NPs) and magnetic nanoparticles (MNPs). The Au-NPs are coated with the target-specific DNA probe which can recognize the target gene, and fluorescein-labeled barcode DNA in a 1:100 probe-to-barcode ratio. The MNPs are coated with the 2nd target-specific DNA probe. After mixing the nanoparticles with the 1st target DNA, the sandwich structure (MNPs-2nd DNA probe/Target DNA/1st DNA probe-Au-NPs-barcode DNA) is formed. A magnetic field is applied to separate the sandwich from the unreacted materials. Then the bio-barcode DNA is released from the Au-NPs. Because the Au-NPs have a large number of barcode DNA per DNA probe binding event, there is substantial amplification. The released barcode DNA is measured by fluorescence. Using this technique, the detection limit of this bio-barcode DNA assay is as low as 2.15 x 10(-16)mol (or 1 ng/mL).