Progress 07/01/05 to 06/30/07
Outputs
The objective of this project is to lower the incidence of foodborne illness and protect the food supply of these United States by rapid detection of the presence of toxins and pathogenic bacteria in food products. Under this research project a new type of wireless biosensor, the magnetostrictive particle (MSP) biosensor, has been designed, fabricated and tested. Testing was conducted in both water and in food products (milk and apple juice). MSP biosensors consist of a rectangular shaped strip of magnetostrictive material that is coated with a bio-molecular recognition element (phage or antibodies) that binds specifically to the target bacteria or spore. The MSP biosensor when subjected to an applied time varying magnetic field oscillates at a characteristic resonance frequency. Binding of target bacteria or spores to the sensor's surface results in a drop in the characteristic resonance frequency. This change in resonance frequency is due to the additional mass of
the bound species and is proportional to the number of bound organisms. Two methods of fabricating the sensor platform have been developed, dicing and microelectronics fabrication. In dicing, ribbons of magnetostrictive materials are cut into sensor platforms using a microelectronics wafer dicing saw. Using this instrument, sensors as small as 35 x 50 x 250 microns have been fabricated. This size sensor is capable of detecting as few as 200 to 500 organisms. The detection limit of the sensor can be improved by making the sensor smaller in size. A sensor of the size 5 x 10 x 50 microns is sensitive enough to detect 1-10 bound organisms. Sensors of this size have been fabricated using microelectronics fabrication methods. Physical vapor deposition is used to co-deposit simultaneously up to three elements to form the rectangular shaped strip sensors. New techniques were developed to handle and measure these very small size magnetostrictive sensors. Both traditional antibodies and
filamentous phage, developed by our team, were used as the bio-molecular recognition elements to construct the MSP biosensors to detect Salmonella typhimurium and Bacillus anthracis. These MSP sensors (35 x 50 x 250 microns in size) were demonstrated to have a detection limit of a few hundreds of cells/ml of analyte. The phage based MSPs have a detection limit better than similar MSP antibody based sensors. In addition to the detection limit experiments, masking experiments were conducted on the MSP sensors. The MSP phage based sensors were shown to be capable of detecting one cell of Salmonella in 105 cells of E. coli. Similarly experiments with Bacillus anthracis Sterne strain spores have been conducted that show detection limits of a few hundreds of spores per ml of liquid. Masking experiments using Bacillus cereus have shown that the MSP biosensors are capable of detecting five Bacillus anthracis spores in a background of 105 cells of Bacillus cereus. MSPs of size 35 x 200 x 1000
microns were tested in food products such as non-fat milk and apple juice. Because larger size sensor platforms were used, the detection limits of these sensors in food products were a few 1000 cells per ml of liquid.
Impacts
Food safety is a national priority which affects every man, woman and child. A crucial part of a prevention strategy to lower the high incidence of foodborne illness is to employ methods that can rapidly detect the presence of toxins and pathogenic bacteria in food products. This research project is focused on the development of RFID (radio-frequency identification) sensor tags for the detection of foodborne bacteria such as Salmonella typhimurium. These RFID sensor tags would be placed on each and every food product sold in the United States, providing all or some of the following resources: automatic inventory, product time and temperature monitoring, and instantaneous traceability. Recently U.S. citizens and farmers have expressed a concern about agro-terrorism. In response to this concern our Center is pursuing detection technologies that can be used to rapidly identify deliberate contaminations of foods with bio-threat agents such as anthrax and Salmonella. The
long range goal is to incorporate these sensors with RFID sensor tags to enable rapid detection of such an attack.
Publications
- R. Guntupalli, Jing Hu, Ramji S. Lakshmanan, Jiehui Wan, Shichu Huang, Hong Yang, James M. Barbaree, T. S. Huang, Bryan A. Chin, Detection of Salmonella typhimurium using polyclonal antibody immobilized magnetostrictive biosensors, Sensors, and Command, Control, Communications, and Intelligence (C3I) Technologies for Homeland Security and Homeland Defense, SPIE Vol. 6201, pp 220-228. May 2006.
- V. A. Petrenko, J. Brigati, J. Sykora, E. Olsen, I.B. Sorokulova, G. Kouzmitcheva, I-H Chen, J. Barbaree, B. Chin and V. Vodyanoy, Landscape Phages and Their Stripped Proteins as Self Assembling Affinity Reagents for Medicine and Technology, Invited Paper, published in European NANO SYSTEMS 2005, Paris, France, pp.81-86, 2006.
- Ramji S Lakshmanan, Jing Hu, Rajesh Guntupalli, Jiehui Wan, Shichu Huang, Hong Yang, Valery A. Petrenko, James M Barbaree, Bryan A. Chin, Detection of Salmonella typhimurium using a Phage-Based Magnetostrictive Sensor, Chemical and Biological Sensing VII, SPIE Vol. 6218, pp. 246-255, May, 2006.
- J. Wan, H. Yang, R. S. Lakshmanan, R. Guntupalli, S. Huang, J. Hu, V. A. Petrenko, and B.A. Chin, Phage-based Magnetostrictive-Acoustic Microbiosensors for Detecting Bacillus anthracis Spores, Micro (MEMS) and Nanotechnologies, SPIE Volume 6223. pp.98-106, May, 2006.
- Vainrub, A., O. Pustovyy, and V. Vodyanoy, Resolution of 90 nm (lambda/5) in an Optical Transmission Microscope with an Annular Condenser, Optics letters 31, pp. 2855-2857 (2006).
- G. Vertelov, W. Gale and A. Simonian, TCP Electrochemical Detection in Chemical Sensors and MEMS, ECS Transactions, (2006) pp.21-34.
- M. H. Hall, P. Shevlin, H. Lu, A. Gichuhi, and C. Shannon, Electron Acceptor-Induced Isomerization of Aryl [6,5] Open Fulleroids to [6,6] Closed Methanofullerenes and the Electrochemical Evaluation of their Free Energy Difference, J. Org. Chem., 2006, 71, pp.3357- 3363.
- O.A. Oyarzabel, The Epidemiology of Campylobacteriosis in Live Broilers (Spanish), Avicultura Profesional 24, (2006) pp.16-17.
- O.A. Oyarzabel, Antimicrobials to Control Campylobacters in Broilers (Spanish), Avicultura Profesional 24, (2006) pp.18-20
- R. Guntupalli, R. S. Lakshmanan, M. L. Johnson, J. Hu, T-S. Huang, J. M. Barbaree, V. J. Vodyanoy, and B. A. Chin, Magnetoelastic Biosensor for the Detection of Salmonella typhimurium in Food Products, Sensing and Instrumentation for Food Quality and Safety, March 2007, Vol. 1, Issue 1, pp.3-10.
- Dong Wei, Omar A. Oyarzabal, Tung-Shi Huang, Shankar Balasubramanian, Srinivas Systa, and Aleksandr L. Simonian, Development of a Surface Plasmon Resonance Biosensor for the Identification of Campylobacter jejuni, Journal of Microbiological Methods, 2007, 69, pp.78-85.
- Viswaprakash Nanduri, Shankar Balasubramanian, Srinivas Sista, Vitaly J. Vodyanoy and Aleksandr L. Simonian, Highly Sensitive Phage-based Biosensor for the Detection of β-galactosidase, Analytica Chimica Acta, Volume 589, pp.6-10 (2006).
- S. Kintzios, I. Marinopoulou, G. Moschopoulou, O. Mangana, K. Nomikou, K. Endo, I. Papanastasiou, A. Simonian, Development of a Novel, Multi-Analyte Biosensor System for Assaying Cell Division: Identification of Cell Proliferation/Death Precursor Events, Biosensors & Bioelectronics, 2006, 21, pp.1365-1373.
- S. Balasubramanian, A. Revzin, A. Simonian, Electrochemical Desorption of Proteins from Gold Electrode Surface, S Electroanalysis, 2006, Invited publication, Volume 18, Issue 19-20, pp. 1885-1892.
- L. Viveros, S. Paliwal, D. McCrae, J. Wild, and A. Simonian, A Fluorescence-based Biosensor for the Detection of Organophosphate Pesticides and Chemical Warfare Agents, Sensors & Actuators B, 2006, 115, pp.150-157.
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Progress 07/01/05 to 06/30/06
Outputs
Safe food is vital to every American's health. Recent incidents involving large-scale accidental contamination and difficulties in tracing/recovering tainted food products indicate the need for new technologies to ensure the safety of our food. The U.S. food system is a complex farm-to-fork continuum, with multiple and ever-changing supply chains to meet consumers' demands. Therefore, protecting the U.S. food supply from accidental or deliberate contamination requires a concerted systems approach. Agro-terrorism can result from an intentionally introduced biological, chemical or nuclear agents that would devastate people, animals or crops. The detection of agro-terrorism agents that may be used to contaminate our food supply and disrupt commerce is therefore a high priority. This project is part of a 10 year effort to develop technologies that will allow rapid detection and tracing of tainted food. Major accomplishments of the project this past year involve the
development of phage as a possible replacement for antibodies as the bio-molecular recognition elements in rapid detection systems for pathogens and toxins. As a demonstration of this technology, phage has been genetically engineered to bind with a pathogenic bacteria (Salmonella typhimurium) and a nonpathogenic strain of spore (Bacillus anthracis Sterne strain). Comparison tests between phage based and antibody based bio-molecular recognition sensors indicate that phage based bio-molecular recognition is 50 to 1000 times more sensitive than antibody based bio-molecular recognition. In addition, phage has a much longer shelf life (5 years) as compared to antibodies (1 year). Both the phage and antibody bio-molecular recognition have been combined with magnetostrictive particle sensors that have been fabricated and investigated under the USDA grants. Using microelectronic manufacturing techniques, rectangular shaped magnetostrictive particles have been fabricated that are 5 x 20 x 100
microns in size. This size of sensor platform theoretically should be able to detect the attachment of a single spore or bacteria. The particles are coated with Ti followed by Au to produce a corrosion resistant and bioactive surface that will accept the deposition of a phage based coating onto the sensor surface. Tests of these biosensors have been conducted in water, buffer solution, milk, apple juice and chicken exudates. A new method of measuring the magnetostrictive particles that enables measurements of many magnetostrictive particles simultaneously has been developed. This new method of measurement also allows tests to be conducted in static as well as flowing liquids. The new measurement technology allows wireless and remote interrogation of the sensors and has been used to measure sensors 100 microns to 5 mm in length. Additionally procedures are being developed to demonstrate a phage based ELISA test kit. This test kit uses phage as an alternate to antibody recognition and
would directly replace currently marketed ELISA type kits in the commercial market.
Impacts
Food safety is a national priority which affects every man, woman and child. A crucial part of a prevention strategy to lower the high incidence of foodborne illness is to employ methods that can rapidly detect the presence of toxins and pathogenic bacteria in food products. This research project is focused on the development of RFID (radio-frequency identification) sensor tags for the detection of foodborne bacteria such as Salmonella typhimurium. These RFID sensor tags would be placed on each and every food product sold in the United States, providing all or some of the following resources: automatic inventory, product time and temperature monitoring, and instantaneous traceability. Recently U.S. citizens and farmers have expressed a concern about agro-terrorism. In response to this concern our Center is pursuing detection technologies that can be used to rapidly identify deliberate contaminations of foods with bio-threat agents such as anthrax and Salmonella. The
long range goal is to incorporate these sensors with RFID sensor tags to enable rapid detection of such an attack.
Publications
- Hong, J.W., Chen, Y., Anderson, N.F., and Quake, S.R., "Molecular Biology on a Microfluidic Chip," Journal of Physics: Condensed Matter, 18, in press (2006)
- Oyarzabal, O. A., N. M. Behnke, M. A. Mozola, W. H. Andrews, E. T. Ryser, and C. W. Donnelly. "Validation of a microwell DNA probe assay for detection of Listeria spp. in foods," Journal of Association of Official Analytical Chemists International. 89:651-668 (2006).
- T. Reeves, Sh. Palival, J. Wild, and A. Simonian. "Orientation Specific Attachment of Organophosphate Hydrolase for Increased Biosensor Performance," Analytica Chimica Acta, submitted (2006).
- Andersen ES, Rosenblad MA, Larsen N, Westergaard JC, Burks J, Wower IK, Wower J, Gorodkin J, Samuelsson T, and Zwieb C. "The tmRDB and SRPDB resources," Nucleic Acids Res. Jan 1:34 (Database issue): D163-8 (2006).
- Pettitt, B. M.; Vainrub, A.; Wong, K.-Y. "DNA saline solutions near surfaces: a few ideas towards design parameters of DNA arrays," NATO Science Series, II: Mathematics, Physics and Chemistry 206 (Ionic Soft Matter: Modern Trends in Theory and Applications), pp. 381-393 Publisher: Springer, CODEN: NSSICD Journal (2006).
- Srinivas Sista, Dong Wei, Omar A. Oyarzabal, and Aleksandr L. Simonian. "Sensitive Surface Plasmon Resonance Biosensor for the Real Time Detection of Campylobacter jejuni," Sensors & Actuators, submitted (2006).
- Shankar Balasubramanian, Iryna Sorokulova, Vitaly Vodyanoy and Aleksandr Simonian. "Lytic Phage as a Specific and Selective Probe for Detection of Staphylococcus aureus - A Surface Plasmon Resonance Spectroscopic Study," Biosensors & Bioelectronics, accepted (2006).
- S. Kintzios, I. Marinopoulou, G. Moschopoulou, O. Mangana, K. Nomikou, K. Endo, I. Papanastasiou, and A. Simonian. "Development of a Novel, Multi-analyte Biosensor System for Assaying Cell Division: Identification of Cell Proliferation/death Precursor Events," Biosensors & Bioelectronics, 21, Pp.1365-1373 (2006).
- L. Viveros, S. Paliwal, D.A. McCrae, J. Wild, and A. Simonian."A Fluorescence-based Biosensor for the Detection of Organophosphate Pesticides and Chemical Warfare Agents," Sensors & Actuators, B, 113, Pp.112-121 (2006).
- A. L. Simonian, T. A. Good, S.-S. Wang, and J. R. Wild. "Nanoparticle-based Optical Biosensors for the Direct Detection of Organophosphate Chemical Warfare Agents and Pesticides," Journal of Analytica Chimica Acta, Vol. 534, Issue 1, Pp. 69-77 (2006).
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