Nanotechnology and Biosensors

Non-Technical Summary:<br/>
A novel biosensor will be developed based on nanomaterials and nanostructures for more rapid and more sensitive detection of pathogenic bacteria and viruses in food and animal samples. This will provide the food and agricultural industry an advanced technology to ensure food safety and control animal diseases.
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Approach:<br/>
The biosensor consists of a sampler, multiple-section microfluidic cartridges, a pumping unit, an impedance detector, a microprocessor, a display, a key panel, and a USB connector. When a food or poultry sample, containing various biological and chemical components with bacteria/virus, is dropped, it is mixed with magnetic nanobeads coated with antibodies/aptamers for several min to get sufficient bioreactions to capture target bacteria/virus. Then, the target bacteria/virus are separated by applying a magnetic field to hold magnetic bio-nanoparticles while washing. During their flowing through a micro/nanofluidics channel, target bacteria/virus are captured by the antibodies/aptamers immobilized on the nanowire/nanoelectrode/ nanochannel. Free nanobeads and others can pass through the channel. The change in impedance, caused by captured target bacteria/virus, is measured and correlated to the concentration of bacteria/virus in a sample. A research prototype of nano-biosensor will be designed, fabricated, and tested. The nano-biosensor will be further optimized, improved and evaluated for its applications in agriculture and foods.
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Progress:<br/>
2012/01 TO 2012/12<br/>
OUTPUTS: By integrating the high efficiency of magnetic nanoparticles-based sample preparation for separation/concentration, the high sensitivity of nanowire/nanoelectrodes, and the high efficacy of flow-through micro/nanofluidics channel into the design, we proposed to develop a nano-biosensor that will meet the required sensitivity, specificity, and speed for screening of pathogenic bacteria and viruses in different foods and poultry samples. The specific objectives of this research are: <br/>(1) Develop new technologies for characterizing fundamental nanoscale processes. Develop a bioseparation method based on magnetic nanoparticles coated with specific antibodies/aptamers to separate target bacteria or virus in a food or poultry sample and concentrate them for detection using a nano-biosensor; <br/>(2) Construct and characterize self-assembled nanostructures. Design and fabricate bio-nanowire/nanotube based micro/nanoelectrodes or quantum dots based fluorescent detector to improve detection sensitivity and reduce assay time; and <br/>(3) Develop devices and systems incorporating microfabrication and nanotechnology. Evaluate the biosensor for rapid detection of L. monocytogenes, S. Typhimurium, E. coli O157:H7 and other pathogenic bacteria in different food samples, and for in-field screening of avian influenza virus in poultry swab samples. <br/>The biosensor consists of a sampler, multiple-section microfluidic cartridges, a pumping unit, an impedance detector, a microprocessor, a display, a key panel, and a USB connector. When a food or poultry sample, containing various biological and chemical components with bacteria/virus, is dropped, it is mixed with magnetic nanobeads coated with antibodies/aptamers for several min to get sufficient bioreactions to capture target bacteria/virus. Then, the target bacteria/viruses are separated by applying a magnetic field to hold magnetic bio-nanoparticles while washing. During their flowing through a micro/nanofluidics channel, target bacteria/viruses are captured by the antibodies/aptamers immobilized on the nanowire/nanoelectrode/ nanochannel. Free nanobeads and others can pass through the channel. The change in impedance, caused by captured target bacteria/virus, is measured and correlated to the concentration of bacteria/virus in a sample. A research prototype of nano-biosensor will be designed, fabricated, and tested. The nano-biosensor will be further optimized, improved and evaluated for its applications in agriculture and foods.
<br/>PARTICIPANTS: Dr. Billy Hargis, Professor of Poultry Health, University of Arkansas; Dr. Steve Tung, Associate Professor of Mechanical Engineering, University of Arkansas; Dr. Walter Bottje, Professor of Poultry Physiology, University of Arkansas; Dr. Huaguang Lu, Research Professor, Animal Diagnostics Laboratory, Penn State University; Dr. Tony Huang, Assistant Professor of Micro/Nanofabrication, Penn State University; Dr. Yibin Ying, Professor of Biosystems Engineering, Zhejiang University, China; Dr. Jianping Wang, Professor of Agricultural Engineering, Zhejiang University, China; Dr. Maohua Wang, Professor of Precision Agriculture, China Agricultural University, China; Dr. Xiwen Luo, Professor of Agricultural Engineering, South China Agricultural University, China; Dr. Ming Liao, Professor of Veterinary, South China Agricultural University, China; Dr. Ronghui Wang, Research Associate of Biology/Biochemistry, University of Arkansas; Lisa Cooney, Program Associate of Microbiology, University of Arkansas; Zach Callaway, PhD student of Biological Engineering, University of Arkansas; Balaji Srinivasan, PhD student of Mechanical Engineering, University of Arkansas; Jacob Lum, MS student of Cell and Molecular Biology, University of Arkansas; Zach Callaway, PhD student of Biological Engineering, University of Arkansas; Yixiang Wang, PhD student of Biological Engineering, University of Arkansas; Luke Brockman, MS student of Biological Engineering, University of Arkansas.
<br/>TARGET AUDIENCES: The poultry and animal industries, and food industries.
<br/>PROJECT MODIFICATIONS: Not relevant to this project.
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Impact:<br/>
Contaminated food is estimated to cause 76 million illnesses, 325,000 serious illnesses resulting in hospitalization, and 5,000 deaths in the United States each year (CDC, 2011). The economic impact of foodborne illness has been estimated as high as $10 billion annually (USDA/ERS, 2002). Current practices for preventing foodborne diseases due to microbial contamination of food products rely upon rapid identification and effective control of specific pathogens from farm to fork. However, conventional culture methods are extremely time-consuming, typically requiring at least 24 h and complicated multi-steps to confirm the analysis. Even current rapid methods such as ELISA and PCR still take 4-8 h to generate only qualitative results and require laboratory setup and skilled personnel. Avian influenza (AI) virus H5N1 has been reported by WHO in more than 46 countries for animal cases and in 15 countries for human cases with 615 people infected and 364 died since 2003. In the US, a recent outbreak of low pathogenic AI in 2001 and 2002 resulted in the depopulation of over 4.5 million chickens and turkeys and had cost the poultry industry approximately $125 million. World Bank estimated that more than 140 million birds had died or been destroyed due to AI H5N1 and losses to the poultry industry are in excess of $10 billion worldwide. A key in controlling the spread of AI is to rapidly detect the disease, and then eradicate infected animals, quarantine and vaccinate animals. The technology for detection of AI H5N1 is mature, but these tests are complex, some are liable to error, and some can be performed safely only in BSL3 facilities. The nanomaterials based biosensors being developed in this project will provide the food industry with more rapid, specific, sensitive and cost-effective method for the detection of pathogens in food products. The quantum dots based fluorescent biosensor is able to detect several cells of L. monocytogenes in a food sample or several hundred cells of Listeria, Salmonella and E. coli O157:H7 simultaneously. The magnetic nanobeads and microfluidics based impedance biosensor can detect AI H5N1 and H5N2 at 103 EID50/ml in a poultry sample. The biosensor developed in this project is rapid, robust and reliable, and suitable for use in the field to detect avian influenza virus, providing the poultry industry with a very needed technology for rapid screening of AIV H5N1, AIV H7 or other infectious diseases related viruses in poultry, such as Newcastle diseases virus and infectious bronchitis virus. This will help the poultry industry be better prepared for poultry diseases, ensure poultry product safety and security, and minimize the testing cost. In general, this research is leading to the development of a portable biosensor instruments for on-line or in-field rapid detection of foodborne pathogens or poultry disease viruses. Therefore, the outcome of this study on biosensors for rapid detection of foodborne pathogens will assist the food industry in their enhancing HACCP programs to ensure food safety and security. The biosensors can also be applied to other areas such as environmental protection and clinical diagnosis.