Progress 10/01/11 to 09/30/16
Outputs
Target Audience:Poultry producers and processors, food producers and processors, analytical laboratories Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided the training in biosensor research to five graduate students, three in biological engineering and two in cell and molecular biology, and the professional development to one postdoctoral research associate in analytical chemistry.The project also provided the interdisciplinary training in biodetection technologiestofour joint-training Ph.D. students in biosystems engineering and food science, three visiting scholars in food science and engineering.During conducting this project, an interdisciplinary and/or multidisciplinary environment was created for researchers from different disciplines, including analytical chemistry, microbiology, food science,and biological engineering, to work togetheras teamsto generate new ideas for developing innovative biosensors with approaches from both science and engineering. How have the results been disseminated to communities of interest?The research results have been presented at American Society of Agricultural and Biological Engineers (ASABE) 2012, 2013, 2014, 2015 and 2016annual international meetings, International Association for Food Protection (IAFP) 2012, 2013, 2014, and 2016 annual meetings, Institute of Food Technologists (IFT) 2015 annual meeting and Institute of Biological Engineering (IBE) 2015 annual meeting.More than twentyarticleshave beenpublished in the journals of Biosensors & Bioelectronics, Sensors and Actuators, Transactions of the ASABE, Sensors, Analyst, Talanta, Journal of Food Protection, Journal of Virological Methods, International Journal of Poultry Science, and others. Invited presentations were given at ASABE 2014, CIGR (International Commission of Agricultural and Biosystems Engineering)2014, the 2015Gordon Research Conference on Nanoscale Science and Engineering for Agriculture and FoodSystems,IFT 2015 Symposium on Challenges and Opportunities for Commercial Biosensors, ICSaid (International Conference on Smart Agriculture Innoventive Development) 2014, 2015, 2016. What do you plan to do during the next reporting period to accomplish the goals?We will continue our research by participation in the renewed NC-1194 project, with reporting under new REEport project ARK02545.
Impacts
What was accomplished under these goals?
This project has shown the nanotechnology-based biosensorscould be applied to the rapid detection ofbacterial pathogens in food and avian influenza in poultry, whichhave greatpotentialas innovative technologies toensure food safetyfor improving human health. For Objective 1, a COMSOL simulation modelwas developed for use in the design and optimization ofa magnetic nanoparticle based bioseparation methods to capture, separate and concentrate target pathogens in food samples. Magnetic separation has been used to pretreat samples to capture, separate and concentrate biological targets in complex samples for further detection. Some simulation models have been developed to predict the magnetic forces applied on magnetic particles (MPs), but very little focus on the complexes of MNPs with biological targets. In this research, a simulation model was developed with the help of Comsol Multiphysics software to describe the magnetic properties of biomolecules coated in magnetic particles to optimize magnetic separation. The model was constructed based on the properties of bacterial cells incorporated with a magnetic separation device. E. coli O157:H7 was used to represent antigenic targets. Experiments in this study were conducted to determine some model parameters and to validate the model. The results showed that the developed model could be used to optimize the magnetic separation in terms of separation efficiency and time based on MP size and surface modification, properties of the biological target, and the magnetic separation device. For one case using E. coli O157:H7 covered with 150 nm MPs, the simulation results indicated that the separation time was 1.3 min traveling 9 mm with a 1.3 T magnetic field strength. The experimental results using the same conditions showed that E. coli O157:H7 had a 91.6% separation efficiency when using 3 min for separation time. Any optimization to increase efficiency and lower time would greatly improve upon current research methods. The model in this research provides a powerful tool in the optimization of magnetic separation involving E. coli with a great potential for other bacteria, viruses, and cells. For Objective 2, first, a quantum dots based biosensing method was developed for simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Listeria monocytogenes, and Salmonella Typhimurium in foods using magnetic nanobeads (MNBs) for separation and quantum dots (QDs) as fluorescence reporters. Streptavidin-coated 25 nm MNBs, conjugated with four corresponding biotin-labeled antibodies, respectively, were used to simultaneously capture and magnetically separate four bacterial pathogens from the food matrix in 45 min. Streptavidin-coated QDs with emission wavelengths of 528, 572, 621, and 668 nm, conjugated with four corresponding biotin-labeled aptamers, were used to label the separated MNB-cell complexes. The fluorescence intensities of all reporting QDs in the MNB-cell-QD complexes were measured simultaneously with a portable spectrometer for quantitation of four different types of bacterial cells. SEM and confocal microscopy were used for characterization of the binding between nanobeads, QDs, and bacterial cells and a simulation model was used to analyze the magnetic separation. Results showed for E. coli O157:H7, S. aureus, L. monocytogenes, and S. Typhimurium, the capture efficiencies of antibodies with 25 nm MNBs were 90.4, 87.5, 92.0, and 92.0%, respectively. The limits of detection for E. coli O157:H7, S. aureus, L. monocytogenes, and S. Typhimurium were 80, 100, 47, and 160 CFU mL-1 in pure culture and 320, 350, 110, and 750 CFU mL-1 in ground beef, respectively. The developed biosensor was capable of simultaneously detecting four bacteria within 2.5 h in a broad range of 101-104 CFU mL-1, showing great potential in multiplex detection of more other foodborne pathogens. Secondly,an aptamer-based bifunctional bio-nanogate, which can selectively respond to target molecules, and control enzymatic reaction for electrochemical measurements, was successfully applied for sensitive, selective, rapid, quantitative and label-free detection of avian influenza viruses (AIV) H5N1. A nanoporous gold film with pore size of ~20 nm was prepared by a metallic corrosion method, and the purity was checked by Energy-Dispersive X-ray Spectroscopy (EDS) study. To improve the performance of the bio-nanogate biosensor, its main analytical parameters were studied and optimized. We demonstrated that the developed bio-nanogate was capable of controlling enzymatic reaction for AIV H5N1 sensing within 1 hour with a detection limit of 2-9 HAU (hemagglutination units), and the enzymatic reaction was able to cause significant current change due to the presence of target AIV. A linear relationship was found in the virus titer range of 2-10 - 22 HAU. No interference was observed from non-target AIV subtypes such as H1N1, H2N2, H4N8 and H7N2. The developed approach could be adopted for sensing of other viruses. For Objective 3,first, a portable fluorescent biosensing system was designed and built and further assessed for in-field detection of three main types of bacterial pathogens that have been associated with the outbreaks of foodborne illness. Using the developed fluorescent nanobiosensor coupled with nanobead-based immunomagnetic separation, we conducted blind tests with the portable device to simultaneously detect E. coli O157:H7, L. monocytogenes, and S. Typhimurium in different food products in three cities selected from three big agricultural provinces in China. Specificity tests showed low interference of this multiplex biosensor from non-targets in food samples. The detection could be done from sampling to results within 60 min. Limits of detections of this method for E. coli O157:H7, L. monocytogenes, and S. Typhimurium were determined to be 102, 103, and 103 CFU/mL in lettuce, shrimp, and ground beef, respectively. Recovery tests were also investigated and this method was evaluated to be accurate comparing with the gold standard culturing method. Therefore, it is feasible for this portable fluorescence biosensing system to be used in rapid and in-field screening of multiple foodborne pathogenic bacteria in foods, such as vegetables, livestock meat, and sea food. And together with fluorescent nanobiosensors, it provides a promising alternative tool to traditional culturing method, or even conventional ELISA and PCR based methods. Secondly,a portable impedance biosensing system was designed to integrate a laptop computer with LabVIEW software, a data acquisition (DAQ) device, a sample delivery system, a micro-flow cell, and a gold interdigitated microelectrode (IDME) into an automatic instrument for rapid detection of AIV H5N1. Streptavidin was first immobilized onto the IDME surface, and then biotin-labeled H5N1-specific aptamer was immobilized on the IDME surface through streptavidin and biotin binding. Samples were delivered through the sample delivery system into the micro-flow cell, and AIV H5N1 was captured by the aptamer on the IDME, resulting in a change in the impedance. A virtual instrument (VI) was programmed with LabVIEW software to provide a platform for sample delivery, impedance measurement, data processing, and control. The audio card of the laptop was used as a function generator, while the DAQ device was used for data communication. The impedance measured by this biosensing system was compared with that measured by a BAS IM6 impedance analyzer, and the error was less than 5%. The results indicated that the developed system could detect AIV H5N1 at a detection limit of 0.84 HAU per 200 mL without interaction with three non-target AIV subtypes (H1N1, H5N2, and H5N3). The system was portable and automatic, and it showed great potential as a diagnostic and epidemiological tool for detection of AIV in the field.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Callaway, Z., Wang, Y.X., B.H Zhang, T.M. Zhang, Z. Zhao, T. Costello, M.F. Slavik, and Y. Li*. 2016. A portable impedance biosensing system for rapid detection of avian influenza virus. Transactions of the ASABE 59(2):421-428.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Karash, S., R. Wang, L. Kelso, H. Lu, T. Huang, and Y. Li*. 2016. Rapid detection of avian influenza virus H5N1 in chicken tracheal samples using an impedance aptasensor with gold nanoparticles for signal amplification. Journal of Virological Methods 236:147-156.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Xu, L.Z., R.H. Wang, L. Kelso, Y.B. Ying, and Y. Li*. 2016. A target-responsive and size-dependent hydrogel aptasensor embedded with QD fluorescent labels for rapid detection of avian influenza virus H5N1. Sensors and Actuators B: Chemical 234:98-108.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Xu, M., R.H. Wang, and Y Li*. 2016. Rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods using an electrochemical immunosensor based on screen-printed interdigitated microelectrode and immunomagnetic separation. Talanta 148:200-208.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Zhang, B.H., R.H. Wang, Y.X. Wang, and Y. Li*. 2016. LabVIEW-based impedance biosensing system for detection of avian influenza virus. International Journal of Agricultural and Biological Engineering 9(4)116-122.
- Type:
Book Chapters
Status:
Published
Year Published:
2016
Citation:
Wang, R., and Y. Li. 2016. Chapter 4. Biosensors for Rapid Detection of Avian Influenza Viruses. P. 61-84, In: Steps Forwards in Diagnosing and Controlling Influenza, M. Baddour (ed). InTech, Rijeka, Croatia. ISBN 978-953-51-2733-8.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Fu, Y.C., Z.Y. Liu, Q.J. Xie, S.Z. Yao, Y. Li*, Y.B. Ying. 2016. Bio-inspired protein-polymer composite adhesive as ultra-highly efficient immobilization matrix for electrochemical biosensing. Presented at Biosensors 2016, May 25-27, 2016, Gothenburg, Sweden. Paper No. P3.132.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Li, Y., Z. Zhao, Z. Callaway, L.Z. Xu, and R. Wang. 2016. Portable biosensors for in-field detection of pathogenic bacteria in foods and mycotoxin in grains. Presented at IBE 2016 annual meeting, April 7-9, 2016, Greenville, SC.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Qiao, Z.H., C.Y. Lei, Y.C. Fu, and Y. Li*. 2016. An immunomagnetic optical sensor for the detection of E. coli O157:H7 based on silver nanoparticles-urease signal amplification. Presented at Biosensors 2016, May 25-27, 2016, Gothenburg, Sweden. Paper No. P1.095.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Wang, H, Q.Q. Hu, R. Wang, Y. Li, and M.T. Kidd. 2016. Rapid detection of Campylobacter jejuni in poultry products using a piezoelectric immunosensor integrated with magnetic immunoseparation. Presented at IAFP 2016 Annual Meeting, July 31-August 3, 2016, St. Louis, MO. Paper No. P1-76.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Wang, L.J., R. Wang, F. Chen, H. Wang, M. Slavik, H. Wei, and Y. Li*. 2016. Development of a sensitive aptamer-based PCR with magnetic immunoseparation for detection of Salmonella Typhimurium in ground turkey. Presented at IAFP 2016 Annual Meeting, July 31-August 3, 2016, St. Louis, MO. Paper No. P1-99.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Wang, R., X.F. Yu, T. Huang, and Y. Li*. 2016. A nanowell-based immunosensor for rapid and sensitive detection of E. coli O157:H7. Presented at IAFP 2016 Annual Meeting, July 31-August 3, 2016, St. Louis, MO. Paper No. P2-46.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Wang, Y.H., M.H. Wang, J.H. Lin, and Y. Li. 2016. A magnetophoretic system for continuous-flow immunoseparation of avian influenza virus. ASABE Paper No. 162461403. Presented at ASABE 2016 Annual International Meeting, July 17-20, 2016, Orlando, FL.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Xu, M., R. Wang, and Y Li*. 2016. A handheld electrochemical biosensor with glucose oxidase-polydopamine based polymetric nanocomposites and Prussian blue modified screen-printed interdigitated microelectrodes for the detection of E. coli O157:H7 in foods. Presented at IAFP 2016 Annual Meeting, July 31-August 3, 2016, St. Louis, MO. Paper No. P2-47.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2016
Citation:
Xu, L.Z., R. Wang, L.C. Kelso, and Y. Li*. 2016. Exploring size-dependent properties of a target-responsive hydrogel aptasensor embedded with QDs for rapid fluorescent detection of viruses. Presented at Biosensors 2016, May 25-27, 2016, Gothenburg, Sweden. Paper No. P1.189.
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Poultry producers and processors, food producers and processors, analytical laboratories Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided the training in biosensor research to four graduate students, three in biological engineering and one in cell and molecular biology, and the professional development to one postdoctoral research associate in analytical chemistry.The project also provided the interdisciplinary training in biodetection technologiesto two joint-training Ph.D. students in biosystems engineering and one visiting scholar in food science.During conducting this project, an interdisciplinary and/or multidisciplinary environment was created for researchers from different disciplines, including analytical chemistry, microbiology, food science,and biological engineering, to work together in the same researchlaboratoryto generate new ideas for developing innovative biosensors with approaches from both science and engineering. How have the results been disseminated to communities of interest?The research results have been presented at American Society of Agricultural and Biological Engineers (ASABE) 2015 annual international meeting, Institute of Food Technologists (IFT) 2015 annual meeting and Institute of Biological Engineering (IBE) 2015 annual meeting. Several articles were published in the journals of Biosensors & Bioelectronics, Transactions of the ASABE, Sensors, and International Journal of Poultry Science. Invited presentations were given at the 2015Gordon Research Conference on Nanoscale Science and Engineering for Agriculture and FoodSystemsand IFT 2015 Symposium on Challenges and Opportunities for Commercial Biosensors. What do you plan to do during the next reporting period to accomplish the goals?First, conduct more experiments to validate the simulation model developed, and then obtain optimal parameters in magnetic separation using the model in order to improve the magnetic nanoparticle based bioseparation method with higher separation efficiency and less separation time. Secondly, design and construct a prototype of an automated biosensing device for in-field or online use to simultaneously detect multiple foodborne pathogens. The prototype instrument should be portable,easy for operation and controlled with a laptop computer. Thirdly, design and fabricate nanopore/nanowire/nanotube modified electrodes for development of new electrochemical biosensors for more sensitive and rapid detection of avian influenza viruses in poultry and pathogens in food products.
Impacts
What was accomplished under these goals?
First, a COMSOL simulation modelwas developed for use in the design and optimization ofa magnetic nanoparticle based bioseparation methods to capture, separate and concentrate target pathogens in food samples. Magnetic separation has been used to pretreat samples to capture, separate and concentrate biological targets in complex samples for further detection. Some simulation models have been developed to predict the magnetic forces applied on magnetic particles (MPs), but very little focus on the complexes of MNPs with biological targets. In this research, a simulation model was developed with the help of Comsol Multiphysics software to describe the magnetic properties of biomolecules coated in magnetic particles to optimize magnetic separation. The model was constructed based on the properties of bacterial cells incorporated with a magnetic separation device. E. coli O157:H7 was used to represent antigenic targets. Experiments in this study were conducted to determine some model parameters and to validate the model. The results showed that the developed model could be used to optimize the magnetic separation in terms of separation efficiency and time based on MP size and surface modification, properties of the biological target, and the magnetic separation device. For one case using E. coli O157:H7 covered with 150 nm MPs, the simulation results indicated that the separation time was 1.3 min traveling 9 mm with a 1.3 T magnetic field strength. The experimental results using the same conditions showed that E. coli O157:H7 had a 91.6% separation efficiency when using 3 min for separation time. Any optimization to increase efficiency and lower time would greatly improve upon current research methods. The model in this research provides a powerful tool in the optimization of magnetic separation involving E. coli with a great potential for other bacteria, viruses, and cells. Secondly, a quantum dots based biosensing method was developed for simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Listeria monocytogenes, and Salmonella Typhimurium in foods using magnetic nanobeads (MNBs) for separation and quantum dots (QDs) as fluorescence reporters. Streptavidin-coated 25 nm MNBs, conjugated with four corresponding biotin-labeled antibodies, respectively, were used to simultaneously capture and magnetically separate four bacterial pathogens from the food matrix in 45 min. Streptavidin-coated QDs with emission wavelengths of 528, 572, 621, and 668 nm, conjugated with four corresponding biotin-labeled aptamers, were used to label the separated MNB-cell complexes. The fluorescence intensities of all reporting QDs in the MNB-cell-QD complexes were measured simultaneously with a portable spectrometer for quantitation of four different types of bacterial cells. SEM and confocal microscopy were used for characterization of the binding between nanobeads, QDs, and bacterial cells and a simulation model was used to analyze the magnetic separation. Results showed for E. coli O157:H7, S. aureus, L. monocytogenes, and S. Typhimurium, the capture efficiencies of antibodies with 25 nm MNBs were 90.4, 87.5, 92.0, and 92.0%, respectively. The limits of detection for E. coli O157:H7, S. aureus, L. monocytogenes, and S. Typhimurium were 80, 100, 47, and 160 CFU mL-1 in pure culture and 320, 350, 110, and 750 CFU mL-1 in ground beef, respectively. The developed biosensor was capable of simultaneously detecting four bacteria within 2.5 h in a broad range of 101-104 CFU mL-1, showing great potential in multiplex detection of more other foodborne pathogens.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Xu, L.Z., Z. Callaway, R. Wang, H. Wang, M.F. Slavik, A. Wang, and Y. Li*. 2015. A fluorescent aptasensor coupled with nanobeads-based immunomagnetic separator for simultaneous detection of four foodborne pathogenic bacteria. Transactions of the ASABE 58(3):891-906. doi:10.13031/trans.58.11089
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Wang, H., Y. Li and M. Slavik. 2014. Rapid detection of Campylobacter jejuni in poultry products using quantum dots and nanobeads based fluorescent immunoassay. International Journal of Poultry Science 13(5):253-259.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Lum, J., R. Wang, B. Haggis, S. Tung, W. Bottje, H. Lu and Y. Li*. 2015. An impedance aptasensor with microfluidic chips for rapid and specific detection of avian influenza H5N1 and H7N2. Sensors 15(8):18565-18578. doi:10.3390/s150818565
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Callaway, Z., R. Wang, and Y. Li*. 2015. Modeling of the bacteria attached with magnetic-nanoparticles for optimization of magnetic separation process. ASABE Paper No. 152190010. Presented at ASABE 2015 Annual International Meeting, July 26-29, 2015, New Orleans, LA. ASABE ITSC Meeting Paper Award.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Xu, L.Z., R.H. Wang, A. Wang and Y. Li*. 2015. Rapid and label-free detection of avian influenza virus H5N1 using a target-responsive hydrogel based fluorescence aptasensor. ASABE Paper No. 152189223. Presented at ASABE 2015 Annual International Meeting, July 26-29, 2015, New Orleans, LA.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Wang, R., L.Z. Xu, and Y. Li*. 2015. Bio-nanogate controlled enzymatic reaction for virus sensing. Biosensors & Bioelectronics. 67:400-407. doi:10.1016/j.bios.2014.08.071
|
Progress 10/01/13 to 09/30/14
Outputs
Target Audience: Poultry industry, instrument industry, Nanotechnology industry Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Two postdoctoral research associates and 1 M.S. and 4 Ph.D. students in biological engineering, molecular biology, analytical chemistry and nanotechnology were involved in this interdisciplinary research. This project provided the great opportunity for them to work as a team to make novel approaches in development of nano-biosensors for detection of viral and bacterial pathogens in agriculture and food. Two postdoctoral research associates and 1 M.S. and 4 Ph.D. students in biological engineering, molecular biology, analytical chemistry and nanotechnology were involved in this interdisciplinary research. This project provided the great opportunity for them to work as a team to make novel approaches in development of nano-biosensors for detection of viral and bacterial pathogens in agriculture and food. How have the results been disseminated to communities of interest? The results have been presented at national and international meetings of professional organizations and societies in engineering, food, and biology, including ASABE, ASV, CIGR, IAFP, IBE, IEEE/Sensors,SRA, Biosensors, for board audience in academia, industries, and regulatory agencies The results have been reported to Tyson Foods, Inc., Ocean Nanotech, LLC, and other food processing companies. The results have been presented at national and international meetings of professional organizations and societies in engineering, food, and biology, including ASABE, ASV, CIGR, IAFP, IBE, IEEE/Sensors,SRA, Biosensors, for board audience in academia, industries, and regulatory agencies The results have been reported to Tyson Foods, Inc., Ocean Nanotech, LLC, and other food processing companies. What do you plan to do during the next reporting period to accomplish the goals? Research prototypes of nano-biosensor systems will be designed, fabricated, and tested for both impedance aptasensor and fluorescent aptasensor. The nano-biosensor systems will be further optimized, improved and evaluated for their applications in rapid detection of viral and bacterial pathogens in agriculture and foods. Research prototypes of nano-biosensor systems will be designed, fabricated, and tested for both impedance aptasensor and fluorescent aptasensor. The nano-biosensor systems will be further optimized, improved and evaluated for their applications in rapid detection of viral and bacterial pathogens in agriculture and foods.
Impacts
What was accomplished under these goals?
Highly pathogenic avian influenza (AI) virus H5N1 has been reported by WHO (March 26, 2012) in more than 46 countries for animal cases and in 15 countries for human cases with 676 people infected and 398 died since 2003. During 2013 to 2014, AI H7N9 was reported not only for poultry but also for human cases with 132 people infected and 37 died. 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 (USDA, 2003). World Bank (2006) 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.Conventional viral culture method is extremely time-consuming, typically requiring several days and complicated multi-steps to confirm the analysis. Relatively rapid methods such as rRT-PCR and ELISA still take more than 8 h to generate only qualitative results and require laboratory setup and skilled personnel. Therefore, rapid screening method is urgently needed by the poultry industry and inspection agencies. In this project, a nano-biosensor was developed by a multidisciplinary team in this project using nanowire and nanoelectrode/nanochannel in combination with magnetic nanoparticles separation. This nano-biosensor method is able to detect highly pathogenic AI H5N1 and low pathogenic AI H5N2 virus at a concentration as low as 10^2 EID50/ml in a poultry swab sample in less than one hour, which is more sensitive and more rapidly than the current real-time RT-PCR and ELISA methods. The combination of biosensor technology with nanotechnology and biotechnology provides very promising future of this new detection method. The results showed that the nano-biosensor method is rapid, robust and reliable, and suitable for use in the field to detect avian influenza virus in poultry, providing the poultry industry and government agencies with a very needed technology for rapid screening of AI H5N1 and other pathogenic subtypes in poultry for more effective surveillance and control of any possible outbreaks of avian influenza. This nano-biosensor technology would help the poultry industry be better prepared for highly pathogenic AI H5N1, ensure poultry product safety and security, and minimize the testing cost. Foodborne pathogens are responsible formillions of cases of diseases each year. There is a growing need for rapid detection of multiple foodborne pathogens. Simultaneous detection enables large scale screening and reduces assay time and cost. In this project, an aptasensor based on magnetic separation with nanobeads and quantum dots (QDs) as fluorescence reporters was developed for rapid, sensitive, specific, quantitative, and simultaneous detection of multiple foodborne pathogens. Salmonella Typhimurium, Escherichia coli O 157: H7, Listeria monocytogenes, and Staphylococcus aureus were used as four model foodborne pathogens. Both streptavidin-conjugated magnetic nanobeads (MNBs) with a diameter of 25 nm and streptavidin-conjugated QDs with emission wavelengths of 528 nm, 572 nm, 621 nm, and 668 nm were incubated with four corresponding biotin-labeled aptamers, respectively. Beads-aptamers conjugates were used to simultaneously capture and magnetically separate four bacterial pathogens in food in 30 min. QDs-aptamer conjugates were used to label separated beads-cell complexes simultaneously. The fluorescence intensities of the reporting QDs on the MNBs-cell-QDs were measured with a portable spectrometer for quantitation of bacterial cells. Results showed that the developed aptasensor was capable of simultaneously detecting four bacteria within 2 h in a broad range with a lower detection limit of 10^1 cfu/ml. This research will lead to the development of a nanotechnology-based portable biosensor instrument for in-field or on-line rapid detection of bacterial pathogens in food. The nano-biosensor can also be applied to the detection of other pathogenic viruses and bacteria in different areas such as environment protection, food safety and quality, and clinical diagnosis in conjunction with available biosensing materials such as specific antibodies, aptamers, enzymes, or DNA/RNA probes.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Li, M., A. Pradhan, W. Wang, and Y. Li. 2013. Prediction of Listeria innocua in fully cooked chicken breast products during post-package thermal treatment. Poultry Science 92:827-835.
Li, M., W. Wang, W. Fang, and Y. Li. 2013. Inhibitory effects of chitosan coating combined with organic acids on Listeria monocytogenes in refrigerated ready-to-eat shrimps. Journal of Food Protection 76(8): 1377-1383.
Wang, W., M. Li, and Y. Li. 2013. Modeling the thermo-ultrasound inactivation of Vibrio parahaemolyticus in shrimps. Journal of Food Protection 76(10): 1712-1718.
Wang, W., M. Li, W. Fang, A. Pradhan and Y. Li. 2013. A predictive model for assessment of decontamination effect of latic acid and chitosan used in combination on Vibrio parahaemolyticus in shrimps. International Journal of Food Microbiology 167(2): 124-130.
Yan, X.F., R. Wang, J.H. Lin, Y.T. Li, M.H. Wang, D. An, P.R. Jiao, M. Liao, Y.D. Yu, and Y. Li. 2013. Impedance immunosensor based on interdigitated array microelectrodes for rapid detection of avian influenza virus subtype H5. Sensor Letters 11(6-7): 1256-1260.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Huang, X.L., Z.D. Xu, Y. Mao, Y.W. Ji, H.Y. Xu, Y.H. Xiong, and Y. Li. 2015. Gold nanoparticle-based dynamic light scattering immunoassay for ultrasensitive detection of Listeria monocytogenes in lettuces. Biosensors & Bioelectronics 66:184-190.
Lin, J.H., M. Li, Y. Li, and Q. Chen. 2015. A high gradient and strength bioseparator with nano-sized immunomagnetic particles for specific separation and efficient concentration of E. coli O157:H7. Journal of Magnetism and Magnetic Materials 378:206-213.
Fu, Y.C., Z. Callaway, J. Lum, R. Wang, J.H. Lin, and Y. Li. 2014. Exploring enzymatic catalysis in ultra-low ion strength media for ion strength increase-based impedance biosensing of virus using a bare interdigitated electrode. Analytical Chemistry 86 (4):1965-1971.
Han, F.F., X. Qi, L.Y. Li, L.J. Bu, Y.C. Fu, Q.J. Xie, M.L. Guo, Y. Li, Y.B. Ying, and S.Z. Yao. 2014. Bio-inspired preparation of fibrin-boned bionanocomposites of biomacromolecules and nanomaterials for biosensing. Advanced Functional Materials 24(31):5011-5018.
8. Hu, Q.Q., X.H. Xu, Z.M. Li, L.Z. Xu, Y. Zhang, J.P. Wang, Y.C. Fu, and Y. Li*. 2014. Detection of acrylamide in potato chips using a fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots. Biosensors & Bioelectronics 54(15):64-71.
Hu, Y.H., C.C. Wang, B. Bai, M.T. Li, R. Wang, and Y. Li. 2014. Rapid detection of Staphylococcus aureus using quantum dots as fluorescent labels. International Journal of Agricultural and Biological Engineering 7(1):77-83.
Wang, H., Y. Li and M. Slavik. 2014. Rapid detection of Campylobacter jejuni in poultry products using quantum dots and nanobeads based fluorescent immunoassay. International Journal of Poultry Science 13(5):253-259.
Wang, H., Y. Li and M. Slavik. 2014. Rapid and simultaneous detection of Salmonella and Campylobacter in poultry samples using quantum dots based fluorescent immunoassay coupled with magnetic immunoseparation. International Journal of Poultry Science 13(11):611-618.
Xu, L.Z., X. Xu, H. Xiong, L.X. Chen and Y. Li*. 2014. Rapid detection of vegetable cooking oils adulterated with inedible used oils using fluorescence quenching method with aqueous CTAB-coated quantum dots. Sensors and Actuators B: Chemical 203:697-704.
Zhou, L, J.P. Wang, D.J. Li, and Y. Li. 2014. An electrochemical aptasensor based on Au nanoparticles dotted graphene modified glassy carbon electrode for label-free detection of bisphenol A in milk samples. Food Chemistry 162:34-40.
- Type:
Journal Articles
Status:
Submitted
Year Published:
2014
Citation:
Wang, L., R. Wang, B.-W. Kong, S. Jin, K.M. Ye, and Y. Li. 2014. A Ca2+-indicator based B cell biosensor for specific and rapid detection of E. coli O157:H7. Scientific Reports (submitted Sept. 9, 2014).
Li, D.J., W.Q. Wang, J.R. Huang, J.P. Wang, and Y. Li. 2014. 3-Mercaptopropionic acid self-assembled monolayer-based piezoelectric flow immunosensor for the direct detection of Escherichia coli O157:H7. Transactions of ASABE (submitted Sept. 20, 2014).
Hu, Q.Q., X.H. Xu, Y.C. Fu, and Y. Li. 2014. A review on rapid methods for detection of acrylamide. Food Control. (submitted Oct. 21, 2014)
Li, Z.M., Y.C. Fu, W.H. Fang, and Y. Li. 2014. Rapid detection of Escherichia coli O157:H7 using an electrochemical impedance immunosensor based on gold screen-printed interdigitated microelectrodes modified with self-assembled monolayers. Sensors and Actuators B: Chemistry. (submitted Nov. 23, 2014)
Xu, L.Z., Z. Callaway, R. Wang, H. Wang, M.F. Slavik, and Y. Li. 2014. A fluorescent aptasensor coupled with nanobeads-based immunomagnetic separator for simultaneous detection of four foodborne pathogenic bacteria. Transactions of the ASABE (submitted Dec. 1, 2014)
Wang, L., R. Wang, X.F. Yu, B.W. Kong, S. Jin, K. Ye, and Y. Li. 2014. A F�rster resonance energy transfer-based B cell biosensor using genetically encoded calcium indicators to rapidly detect Escherichia coli O157:H7. Applied and Environmental Microbiology (submitted Dec. 23, 2014)
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
Callaway, Z., Y. Fu, R. Wang, J. Lum, and Y. Li. 2013. Modeling the electro-magnetic properties of avian influenza virus in a flow cell with an interdigitated nanoelectrode using Comsol. ASABE 2013 Annual International Meeting, July 21-24, 2013, Kansas City, MO. ASABE Paper No. 131620936.
Wang, R., Y. Fu, and Y. Li. 2013. Impedance immunosensor with disposable printed interdigitated electrodes for rapid detection of E. coli O157:H7 in Foods. ASABE 2013 Annual International Meeting, July 21-24, 2013, Kansas City, MO. ASABE Paper No. 131620230.
Wang, H., Li, Y., and M. Slavik. 2013. Rapid detection of Campylobacter jejuni in poultry products using quantum dots and nanobeads based fluorescent immunoassay. Presented at IAFP 2013 Annual Meeting, July 28-31, 2013, Charlotte, NC. Paper No. P2-108.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Abdullah, S.S., R. Wang, and Y. Li. 2014. Aptamer and microelectrode based impedance assay for detection of H5N1 influenza virus. Presented at ASV 2014 33rd Annual Meeting, June 21-23, 2014, Fort Collins, CO. Paper No 29.
Callaway, Z., R. Wang, and Y. Li. 2014. Modeling the electromagnetic properties of E. coli cells with different components of biological immobilization components on a screen-printed interdigitated microelectrode using Comsol. Presented at ASABE 2014 Annual International Meeting, July 13-16, 2014, Montreal, Canada. ASABE Paper No. 1897698.
Fu, Y.C., F.F. Han, Q.J. Xie, and Y. Li. 2014. Bio-inspired preparation of fibrin-boned bionanocomposites of biomolecules and nanomaterials for electrochemical biosensing. Presented at the 24th World Congress on Biosensors, May 27-30, 2014, Melbourne, Australia. Paper No. P1.171.
Fu, Y.C., X. Qi, and Y. Li. 2014. Detecting glucose in honey using polymer-based enzyme biosensors. Presented at 18th World Congress of CIGR, September 16-19, 2014, Beijing, China. Paper No. 71-002.
Hu, Q.Q., X.H. Xu, Y.C. Fu, and Y. Li. 2014. Colorimetric detection of acrylamide based on its metabolite-induced inhibition of hybridization of gold nanoparticles-conjugated DNA enhanced by enzymatic catalysis. Presented at the 24th World Congress on Biosensors, May 27-30, 2014, Melbourne, Australia. Paper No. P3.111.
Li, Z.M., Y.C. Fu, and Y. Li. 2014. An electrochemical impedance immunosensor for the detection of Escherichia coli O157:H7 with lectin based signal amplification. Presented at the 24th World Congress on Biosensors, May 27-30, 2014, Melbourne, Australia. Paper No. P1.169.
Li, Z.M., Y. Fu, and Y. Li. Self-assembled monolayers-based impedance immunosensor for rapid detection of Escherichia Coli O157:H7 using screen-printed interdigitated microelectrodes. Presented at CIGR2014 International Meeting, September 15-19, 2014, Beijing, China. Paper No. 2-14-0962.
Lin, J.H., R. Wang, P.X. Jiao, Y.T. Li, X.H. Wen, Y. Li, M. Liao, and M.H. Wang. 2014. An improved impedance biosensor based on interdigitated array microelectrode for rapid detection of avian influenza virus. Presented at the 24th World Congress on Biosensors, May 27-30, 2014, Melbourne, Australia. Paper No. P2.020.
Wang, H., Y. Li, and M.F. Slavik. 2014. Rapid and simultaneous detection of Campylobacter and Salmonella in poultry samples using magnetic nanobeads and quantum dots based fluorescent immunosensor. Presented at IAFP 2014 Annual Meeting, August 3-6, 2014, Indianapolis, IN. Paper No. P2-92.
Wang, R., and Y. Li. 2014. Bio-nanogate controlled enzymatic reaction for virus sensing. Presented at the 24th World Congress on Biosensors, May 27-30, 2014, Melbourne, Australia. Paper No. P1.141.
Wang, R., L. Wang, X.F. Yu, B.W. Kong, and Y. Li. 2014. Fluorescent Ca2+ indicator based B Cells biosensor for rapid detection of E. coli O157:H7 in foods. Presented at IAFP 2014 Annual Meeting, August 3-6, 2014, Indianapolis, IN. Paper No. P2-116.
Wang, Y.X., B.H. Zhang, R. Wang, S.S. Abdullah, and Y. Li. 2014. A portable impedance biosensing system based on a laptop with LabVIEW for detection of avian influenza virus. Presented at ASABE 2014 Annual International Meeting, July 13-16, 2014, Montreal, Canada. ASABE Paper No. 1897866.
Xu, M., R. Wang, and Y. Li. 2014. Screen-printed electrode based aptasensor for rapid detection of E. coli O157:H7 in foods. Presented at IAFP 2014 Annual Meeting, August 3-6, 2014, Indianapolis, IN. Paper No. P2-149.
Xu, L.Z., Z. Callaway, R. Wang, and Y. Li. 2014. A fluorescent aptasensor coupled with nanobeads-based immunomagnetic separation for simultaneous detection of four foodborne pathogens. Presented at ASABE 2014 Annual International Meeting, July 13-16, 2014, Montreal, Canada. ASABE Paper No. 1895935.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2014
Citation:
Lum, J. 2014.Impedance biosensor for the rapid detection of viral and bacterial pathogens using avian influenza virus subtypes H5N1 and H7N2 and Escherichia coli O157:H7 as model targets. Ph.D. Dissertation, University of Arkansas, Fayetteville, AR.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2014
Citation:
Wang, Y. 2014. A portable and automated biosensor for detection of avian influenza viruses. M.S. Thesis, University of Arkansas, Fayetteville, AR.
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience: Poultry and food industries Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? One M.S. student graduated after his two years research on QCM biosensor for rapid detection of avian influenza virus. One Ph.D. student has completed his disertation on impedance aptasensors fordetection of avian influenza H5N1 and H7, submitted two manuscripts to refeered scientific journals for publictions,and is ready for grdaution. Fivegraduate students have been working on nanotechnology-based biosensors for their Ph.D. or M.S. degrees in biological engineering, cell and molecular biology, or microelectronics and photonics. How have the results been disseminated to communities of interest? We presented the research results with two papers atASABE (American Society of Agricultural and Biological Engineers) 2013 international annual meeting, one paper at IAFP (International Association of Food Protection) 2013 international meeting, three papers atIBE (Institute of Biological Engineering) 2013 annual meeting, and three papers atIEEE(Instiute of Electrical and Electronics Engineers) Sensors 2013 annual meeting. What do you plan to do during the next reporting period to accomplish the goals? During the next year, we plan to continue conducting experiments on the magnetic nanoparticle-based capture and separation of multiple pathogenic bacteria in food and viruses in poultry to achieve higher capture/separation efficiency in shorter time. Also,we will continue investigating the characteristics of nanoparticles, nanopores and nanowires when they are used in the biosensors, and then start the design and fabrication of desired nanomaterials. At the same time, we will design the prototype instrument for nanotechnology-based biosensor system, including both the separation and detection componentswith the hardware and software.
Impacts
What was accomplished under these goals?
Based on our previous experiences, we conducted some experiments to prove the concept, as stated as the follows,of nanotechnology-based biosensors for rapid detection of bacteria and virus. 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. During the past year, for Objective (1), magnatic nanoparticles were modified with either specific antibodies or aptamers, and successfully used to capture and separate either avian influenza virus H5N1 and H7 in poultry swab samples or multiple pathogenic bacteria including E. coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, and Campylobacter jejuni in food products.For Objective (2), nanoparticles, nanowires, nanotubes and nanopores were investigated for their charateristics for use in the biosensorsto improve the sensitivity and specificity indetecting bacteria and viruses. New quantun dots were also tested for simultaneous detection of multiple bacteria or viruses. Nanopore-based electrochemical biosensor showed very encouraging results for ultra-sensitive detection of avian influenza.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Brockman, L., R. Wang, J. Lum, and Y. Li. 2013. A QCM aptasensor for rapid and specific detection of avian influenza virus. Open Journal of Applied Biosensor 2(4):97-103.
Chen, P., Y. Li, T. Cui, and R. Ruan.2013. Nanoparticles based sensors for rapid detection of foodborne pathogens. International Journal of Agricultural and Biological Engineering 6(1):1-7.
Hu, Q.Q., X.H. Xu, Z.M. Li, L.Z. Xu, Y. Zhang, J.P. Wang, Y.C. Fu, and Y. Li. 2013. Detection of acrylamide in potato chips using a fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots. Biosensors & Bioelectronics 54(15):64-71.
Wang, R., and Y. Li. 2013. Hydrogel based QCM aptasensor for detection of avian influenza. Biosensors & Bioelectronics 42:148-155.
Wang, R., J. Zhao, T. Jiang, Y.M. Kwon, H. Lu, P. Jiao, M. Liao, and Y. Li. 2013. Selection and characterization of DNA aptamers for use in detection of avian influenza H5N1. Journal of Virological Methods 198: 362-369.
Xu, X., X. Liu, Y. Ying, and Y. Li. 2013. A simple and rapid optical biosensor for detection of aflatoxin B1 based on competitive dispersion of gold nanorods. Biosensors & Bioelectronics 47C:361-367.
Zhou, L., Wang, J.P., L. Gai, D. Li, and Y Li. 2013. An amperometric sensor based on ionic liquid and carbon nanotube modified composite electrode for the determination of nitrite in milk. Sensors and Actuators B: Chemical 181:65-70.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Hu, Q., X.H. Xu, M.Z. Li, Y. Zhang, and Y. Li. 2013. Rapid detection of acrylamide in food using a fluorescent sensing method based on functional CdSe/ZnS quantum dots. The Proceedings of IEEE Sensors 2013 Conference, November 3-6, 2013, Baltimore, MD. Paper No. 7198.
Xu, L.Z., X.H. Xu, H. Xiong, L.X. Chen, and Y. Li. 2013. Identification of adulterated vegetable cooking oils using fluorescence quenching method with aqueous CTAB-Coated CdSe/ZnS quantum dots as probes. The Proceedings of IEEE Sensors 2013 Conference, November 3-6, 2013, Baltimore, MD. Paper No. 7114.
Zhang, B.H., R. Wang, Y. Wang, and Y. Li. 2013. A portable impedance biosensor for detection of multiple avian influenza viruses. The Proceedings of IEEE Sensors 2013 Conference, November 3-6, 2013, Baltimore, MD. Paper No. 7505.
- Type:
Theses/Dissertations
Status:
Other
Year Published:
2013
Citation:
Brockman, L. 2013. QCM aptasensor for rapid and specific detection of avian influenza virus. M.S. Thesis, University of Arkansas, Fayetteville, AR.
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Progress 01/01/12 to 12/31/12
Outputs
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: (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; (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 (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. 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. 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. TARGET AUDIENCES: The poultry and animal industries, and food industries. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
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 10^3 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.
Publications
- Kanayeva, D., R. Wang, D. Rhoads, G. Erf, M. Slavik, S. Tung, and Y. Li. 2012. Efficient separation and sensitive detection of Listeria monocytogenes using an impedance immunosensor based on magnetic nanoparticles, microfluidics and interdigitated microelectrode. Journal of Food Protection 75(11):1951-1959.
- Lum, J., R. Wang, K. Lassiter, B. Srinivasan, D. Abi-Ghanem, L. Berghman, B. Hargis, S. Tung, H. Lu, and Y. Li. 2012. Rapid detection of avian influenza H5N1 virus using impedance measurement of immuno-reaction coupled with RBC amplification. Biosensors & Bioelectronics 38(1):67-73.
- Xu, X., Y. Ying, and Y. Li. 2012. One-step and label-free detection of alpha-fetoprotein based on aggregation of gold nanorods. Sensors and Actuators B: Chemical 175:194-200.
- Zhou, L., D. Li, L. Gai, J. Wang, and Y. Li. 2012. Electrochemical aptasensor for the detection of tetracycline with multi-walled carbon nanotubes amplification. Sensors and Actuators B: Chemical 162:201-208.
- Bai, H., R. Wang, B. Hargis, H. Lu and Y. Li. 2012. A SPR aptasensor for detection of avian influenza virus H5N1. Sensors 12:12506-12518.
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