PORTABLE MAGNETIC PARTICLE SPECTROSCOPY (MPS) PLATFORM FOR ON-FIELD DETECTION OF INFLUENZA A VIRUS AND MYCOPLASMA HYOPNEUMONIAE

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: AFRI COMPETITIVE GRANT
• Reporting Frequency: Annual
• Accession No.: 1023707
• Grant No.: 2020-67021-31956
• Cumulative Award Amt.: $414,999.00
• Proposal No.: 2019-06437
• Project Start Date: Aug 1, 2020
• Project End Date: Jan 31, 2024
• Grant Year: 2020
• Program Code: [A1521]- Agricultural Engineering

Recipient Organization
UNIVERSITY OF MINNESOTA
200 OAK ST SE
MINNEAPOLIS,MN 55455-2009

Performing Department
Electrical and Computer Eng

Non Technical Summary
Porcine respiratory disease poses a major challenge to both veterinarians and pork producers. Conventional methods for identification of pathogens causing porcine respiratory disease are time consuming and require specialized laboratory facilities. It also involves multiple tests to detect different pathogens. MPS technology, proposed in this project, can be used to develop an accurate, affordable, fast and easy-to-use test for pathogen detection, including simultaneous detection of multiple pathogens in a single test. Using the MPS platform as a portable in-farm testing kit, would bring infectious disease diagnosis in swine production systems to a new level. At the successful completion of this project, we would have an MPS system that will enable early rapid detection of two common porcine respiratory pathogens, allowing implementation of quick and effective prevention, and control strategies for these respiratory infections in swine. One could also envision the potential for extending this technology to other swine diseases, and to infections in other animal species. The portability of this detection device coupled with a smartphone interface will allow testing in remote areas and under field settings, such as in a farm or a production system. By transmitting test results collected from distant locations to centrally located data analysis units, veterinarians located away from farms can have real-time epidemiological data at their fingertips. This would also significantly reduce the costs for monitoring infections at the regional level. These possibilities would alter current paradigms for disease surveillance and enhance the ability to evaluate and monitor disease control protocols in real-time.

Animal Health Component

100%

Research Effort Categories

Basic

0%

Applied

100%

Developmental

0%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
311	3510	1101	100%

Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3510 - Swine, live animal;

Field Of Science
1101 - Virology;

Keywords

magnetic particle spectroscopy

porcine respiratory disease

multiplexed diagnostics

point-of-care

Goals / Objectives
Goal 1: Development of a handheld MPS bioassay device with user interface.Circuit board design and assembly.User interface design including application and software.Handheld device structure design.Goal 2: Application of MPS handheld platform and bench-top system for the detection of IAV andMHYOP.Determination of optimum concentration of capture antibodies for IAV andMHYOP.Validation of MPS platform for the multiplexed detection of IAV andMHYOP.Goal 3: Validation of MPS platform for monitoringofIAV andMHYOP using field samples.Evaluation of the MPS platform for IAV andMHYOP diagnosis using field samples.Goal 4:Application of MPS platform for detectingSARS-CoV-2.Validation and evaluation of the MPS platform for SARS-CoV-2diagnosis on purifed and clinical samples.

Project Methods
Goal 1:Development of a handheld MPS bioassay device with user interfaceIn the first stage of this project, we will focus on the development of handheld MPS platform, which consists of a portable MPS device and user interface application. Each test kit containing 100 µL (contains100µgFe3O4MNP) surface functionalized MNPs (MNPs conjugated with capture antibodies targeting on specific biomarker) along with MPS platform allows forrapid, sensitive, and wash-free detection of pathogens. It's worth mentioning that we have already developed the first generation of MPS portable device with user application that is available through the Google Play store (see the team link:https://magicoil.ece.umn.edu/). Thus, in aim 1, the main objective is to further optimize the MPS platformGoal 2:Application of MPS handheld platform and bench-top system for the detection of IAV and MHYOP.Single-pathogen detection will first be conducted to determine the sensitivity and specificity for each of the target pathogens and to optimize the concentrations of capture antibodies in the immunoassays. Subsequently, the MNPs will be functionalized with different capture antibodies specific to each of the target pathogens. Stock cultures ofIAV and MHYOPwill then be mixed together for the multiplexed detection in pure culture. The immunoassays will be further optimized based on the results of the detection. The sensitivity and selectivity will be determined for each of the target pathogens in the multiplexed immunoassay and compared with the results from ELISA and RT-PCR.Goal 3:Validation of MPS platform for monitoring ofIAV and MHYOPusing field samplesTo evaluate the influence of biological sample matrix on the performance of the magnetic immunoassay, known quantities of pathogen will be spiked in different types of pig biological fluids for the multiplexed detection. The detection of real samples from the farm without spiked specimens will then be performed with the sensitivity and selectivity compared to RT-PCR.At the end of the project, it is expected that a portable handheld MPS platform integrated with 100 µL MNP test kit will be developed for the rapid, onsite, and multiplexed detection ofporcine respiratorypathogens. Ultimately, this technology could be transformed into commercially available products for the detection, surveillance, and control of theporcine respiratorypathogens, which could largely benefit thepork industrynot only in US, but throughout the world.Goal 4: Application of MPS platform for detecting SARS-CoV-2At the end of this project, we will extend the application of our MPS platform for detecting SARS-CoV-2 biomarkers.

Progress 08/01/20 to 01/31/24

Outputs
Target Audience:The direct impact of this project will be on 1) hospitals, clinics, doctors, patients that require fast and on-site COVID-19 diagnosis (Goal 4); 2) and veterinarians and pork producers who require field tests for porcine respiratory diseases (Goals 2&3). Changes/Problems:Due to the COVID-19 lock down, the mycoplasma and swine work was shut down (Goal 2 & 3). Thus, we continued our work to build a diagnostic assay for SARS-CoV-2 detection (Goal 4). In the first award year, one major challenge that severely delayed our research progress came from the magnetic nanoparticle supplier, who could not provide us with stable and high-performance nanoparticles for MPS-based bioassays. This challenge was overcome with two solutions: 1) instituting a monthly contract with the company to ensure regular supply of magnetic nanoparticles; 2) expanding our vendor pool to search for other sources for magnetic nanoparticle as backup. What opportunities for training and professional development has the project provided? During the whole period (2020-2024), three senior design teams from the ECE department were involved in the design and realization of the handheld MPS device. The senior design teams worked on designing analog conditioning circuit, firmware implementation, and successful designing of user interface for desktop and mobile devices. Spring 2020: Sesnior design team of Five undergrad students: Owen Roe, Li-Sha Lim, Jacob Cantrell, Grant Udstrand, Joe Bajda worked on the first version implementation of the MPS handheld device. Spring 2022: A team of Three undergrad students:Arturo di Girolamo, Prince Jappah, and Connor Templeton worked on improving the device sensitivity by realization of lock-in mixer implementation. A mobile application for communication with the handheld device was also developed. Spring 2024: A team of Five undergrad members:Aaron Knoll, Austin Friesenhan, Maddie Rowe, Megan Adamek, Mohamad Ibrahim worked towards improving the SNR performance by improving circuit board layout and better ADC implementation. The team also delivered Desktop user interface for allowing user communication. Apart from the involvement of the senior design teams, we also involved the following students for MPS research: Allison Benjamin: High-school student was involved to this project in Summer 2020 to perform qualitative analysis, assessing the impact on COVID-19 on Africal healthcare workers. Rachel Weidmayer: DVM student at UMN,worked on characterizing the Mycoplasma hypopneumoniae reagents that will be used for the MPS assay in Summer 2021. Professional develoment oportunities were provided for two postdocs: Kai Wu, and Venkatraman D Krishna by way of invited talks and poster presentations. Similar opportunities for the two grad students: Vinit Kumar Chugh, and Shuang Liang were also provided through conference attendence and poster presentations. How have the results been disseminated to communities of interest?Apart from the results being published in the scientific journals and the conference poster presentations mentioned in the products section, the results of the research have also been dessiminated by way of News media coverage as well as Invited talks as mentioned below: News Media: IEEE Spectrum:https://spectrum.ieee.org/a-better-faster-cheaper-test-for-covid19 KSTP-TV:https://kstp.com/health-medical/u-of-m-researchers-develop-new-portable-coronavirus-testing-device/ EquiManagement:https://equimanagement.com/news/umn-vet-school-researchers-working-to-develop-covid-19-breathalyzer-test/ UMN College of Veterinary Medicine:https://vetmed.umn.edu/news/newly-funded-developing-widely-accessible-covid-19-breathalyzer-test UMN Office of the Vice President for Research:https://research.umn.edu/news/handheld-device-quickly-detect-covid-19 UMN ECE:https://wayback.archive-it.org/8236/20210527220222/https:/ece.umn.edu/multidisciplinary-team-work-on-development-of-handheld-covid-19-diagnostic-device/ Combat Corona Virus at ECE:https://coronavirus.ece.umn.edu/ Group's research website at ECE:https://magicoil.ece.umn.edu/ Invited Talks: The 13th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, June 14-17, 2022. London, UK. Presenter: Jian-Ping Wang. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Washfree Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. Department of Electrical and Computer Engineering, Texas Tech University. Feb 7, 2022. Lubbock, TX, U.S. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Department of Electrical and Computer Engineering, Texas A&M University. Jan 28, 2022. Virtual. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Department of Electrical and Computer Engineering, University of Minnesota. Sep 16, 2022. Virtual. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Department of Electrical and Computer Engineering, Texas Tech University. Oct 7, 2022. Lubbock, TX, USA. Presenter: Kai Wu. Title: Magnetic Particle Spectroscopy Point-of-Care Device for Disease Diagnosis. The National Institute of Standards and Technology (NIST). Apr 20,2023. Virtual. Presenter: Kai Wu. Title: Magnetic Particle Spectroscopy Point-of-Care Device for Disease Diagnosis. 14th International Conference on the Scientific and Clinical Applications of Magnetic Carriers,June 17-21, 2024,Barcelona, Spain. Presenter: Jian-Ping Wang. Title: Magnetic Particle Spectroscopy (MPS)-based Bioassays. IEEE International Magnetics Conference, INTERMAG 2024, May 5-10, 2024,Rio de Janeiro, Brazil. Presenter: Jian-Ping Wang. Title:Magnetic Particle Spectroscopy (MPS)-based Bioassays. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In the 4 years of of this award, the following goals have been accomplished: Goal 1: Development of a handheld MPS bioassay device with user interface. Two handheld prototypeshave been built with sufficient surface-based assay sensitivity needed for real-world disease diagnostic applications. Sensitivity for volumetric assay implementation still lacks this goal by a sensitivity factor of 40x. User interface (mobile application) has also been developed to enable user communication. Handheld device structure and geometry are also complete. Goal 2: Application of MPS handheld platform and bench-top system for detection of IAV and MHYOP. This goal was partially achived. We were succesfully able to functionalize IAV capture antibodies on the MNPs needed for IAV protein assay implementation. However, the functionalization of MHYOP antibodies of the MNPs was unsuccessfull. This limited our ability towards successfull implementation of the MHYOP assay implementation. Due to the unsuccessful congugation of MHYOP antibodies, the multiplexes assay implementation also could not be achieved. Goal 3: Validation of MPS platform for monitoring of IAV and MHYOP using field samples. IAV assay was implemented with purified N protein as well as spiked whole virus sample achieving a sensitivity of 128 pM and 103 TCID50/ml respectively. MHYOP assays were not run due to the limitation detailed above. Goal 4: Application of MPS platform for detecting SARS-CoV-2. We successfully achieved 5-minute volumetric assay implementation for SARS-CoV-2 (purified) Spike-protein with LOD of 5nM. Surface-based MPS assay was also developed for improving sensitivity results and achieving a LOD of 104 pfu/ml (ELISA LOD: 103 pfu/ml, Lateral flow assay LOD: 102 pfu/ml) with wholevirus spiked serum samples.

Publications

• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: 2020 Design of Medical Devices Conference. Minneapolis, Minnesota, USA. April 69, 2020. Title: Magnetic Particle Spectroscopy-Based Handheld Device for Wash-Free, Easy-to-Use, and Solution-Phase Immunoassay Applications.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Chugh, V.K., Di Girolamo, A., Liu, J., Saha, R., Su, D., Krishna, V.D., Nair, A., Davies, W., Wang, Y.A. and Cheeran, M.C., 2021. A Portable Magnetic Particle Spectrometer for Future Rapid and Wash-Free Bioassays. ACS Applied Materials & Interfaces, 13(7), pp.7966-7976.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Liu, J., Saha, R., Peng, C., Su, D., Wang, Y.A. and Wang, J.P., 2021. Investigation of Commercial Iron Oxide Nanoparticles: Structural and Magnetic Property Characterization. ACS omega, 6(9), pp.6274-6283.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Liu, J., Saha, R., Ma, B., Su, D., Chugh, V.K. and Wang, J.P., 2021. Stable and Monodisperse Iron Nitride Nanoparticle Suspension for Magnetic Diagnosis and Treatment: Development of Synthesis and Surface Functionalization Strategies. ACS Applied Nano Materials, 4(5), pp.4409-4418.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Yari, Parsa, Bahareh Rezaei, Clifton Dey, Vinit Kumar Chugh, Naga Venkata Ravi Kumar Veerla, Jian-Ping Wang, and Kai Wu. "Magnetic particle spectroscopy for point-of-care: a review on recent advances." Sensors 23, no. 9 (2023): 4411.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Shuang Liang, Parsa Yari, Kai Wu, and Jian-Ping Wang. "A method for multiplexed and volumetric-based magnetic particle spectroscopy bioassay: mathematical study." Journal of Physics D: Applied Physics 56, no. 31 (2023): 315001.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Chugh, V.K., Wu, K., Krishna, V.D., di Girolamo, A., Bloom, R.P., Wang, A. (Y.Q.), Saha, R., Liang, S., Cheeran, M.C., & Wang, J.P. (2021) Magnetic Particle Spectroscopy with One-stage Lock-in Implementation for Magnetic Bioassays with Improved Sensitivities. Journal of Physical Chemistry C, 2021, 125, 31, 1722117231.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Chugh, V.K., Krishna, V.D., di Girolamo, A., Wang, A. (Y.Q.), Saha, R., Liang, S., Cheeran, M.C., & Wang, J.P. (2021) One-step, Wash-free, Nanoparticle Clusteringbased Magnetic Particle Spectroscopy Bioassay Method for Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. ACS Applied Materials & Interfaces, 2021, 13, 37, 4413644146.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Wu, K., Liu, J., Chugh, V.K., Liang, S., Saha, R., Krishna, V.D., Cheeran, M.C., & Wang, J.P. (2022) Magnetic Nanoparticles and Magnetic Particle Spectroscopy-based Bioassays: A 15-year Recap. Nano Futures, 6, 022001.
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Wu, K., Chugh, V.K., Krishna, V.D., Wang, Y., Gordon, T.D., Cheeran, M.C., & Wang, J.P. (2022) Five-Minute Magnetic Nanoparticle Spectroscopy-Based Bioassay for Ultrafast Detection of SARS-CoV-2 Spike Protein. ACS Applied Nano Materials, 5(12), pp.17503-17507.
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: The 15th Joint MMM-INTERMAG Conference (2022 Joint), January 10-14, 2022. New Orleans, U.S. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Arturo di Girolamo, Venkatramana D. Krishna, Kai Wu, Maxim CJ Cheeran, and Jian-Ping Wang. "Frequency and amplitude optimizations for magnetic particle spectroscopy applications." The Journal of Physical Chemistry C 127, no. 1 (2022): 450-460.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Su, Diqing, Kai Wu, Karthik Srinivasan, Zohreh Nemati, Reza Zamani, Vinit Chugh, Renata Saha et al. "Ultra?flexible giant magnetoresistance biosensors for lab?on?a?needle biosensing." Advanced Materials Interfaces 10, no. 7 (2023): 2201417.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Shuang Liang, Denis Tonini, Renata Saha, Jinming Liu, Parsa Yari, Venkatramana D. Krishna, Maxim C-J. Cheeran, Kai Wu, and Jian-Ping Wang. "Static and Dynamic Magnetization Responses of Self-Assembled Magnetic Nanoparticle Chains." The Journal of Physical Chemistry C 127, no. 37 (2023): 18494-18505.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Yari, Parsa, Shuang Liang, Vinit Kumar Chugh, Bahareh Rezaei, Shahriar Mostufa, Venkatramana Divana Krishna, Renata Saha et al. "Nanomaterial-Based Biosensors for SARS-CoV-2 and Future Epidemics." Analytical Chemistry 95, no. 42 (2023): 15419-15449.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: IEM Annual Conference, September 26, 2023. Minneapolis, U.S. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: The 68th Annual Conference on Magnetism and Magnetic Materials (MMM 2023), October 30 - November 3, 2023. Dallas, U.S. Title: Systematic Analysis of Parameters Affecting Colorization Performance for Magnetic Particle Spectroscopy (MPS) Applications.
• Type: Book Chapters Status: Published Year Published: 2024 Citation: Vinit Kumar Chugh, Kai Wu, Jian-Ping Wang. Magnetic particle spectroscopy (MPS) biosensor. Book  Magnetic Nanoparticles in Nanomedicine, Elsevier (2024).
• Type: Conference Papers and Presentations Status: Other Year Published: 2020 Citation: MMM 2020 Virtual Conference, November 6, 2020. Title: Portable Magnetic Particle Spectroscopy (MPS) Device for Future Rapid, One-step and Wash-free Bioassays.

Progress 08/01/23 to 01/31/24

Outputs
Target Audience:The direct impact of this project will be on 1) hospitals, clinics, doctors, patients that require fast and on-site COVID-19 diagnosis (Goal 4); 2) and veterinarians and pork producers who require field tests for porcine respiratory diseases (Goals 2&3). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have involved five senior students, Aaron Knoll, Austin Friesenhan, Maddie Rowe, Megan Adamek, and Mohamad Ibrahim in this project from Spring 2024. This senior design team worked towards SNR improvement for the handheld MPS prototypes. They also contributed towards creating a Desktop interface for user communication. How have the results been disseminated to communities of interest?The results were dissimated to the communities of interest through two invited talks as mentioned below: 14th International Conference on the Scientific and Clinical Applications of Magnetic Carriers,June 17-21, 2024,Barcelona, Spain. Presenter: Jian-Ping Wang. Title: Magnetic Particle Spectroscopy (MPS)-based Bioassays. IEEE International Magnetics Conference, INTERMAG 2024, May 5-10, 2024,Rio de Janeiro, Brazil. Presenter: Jian-Ping Wang. Title:Magnetic Particle Spectroscopy (MPS)-based Bioassays. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Work focused towards completion of goals-3 and 4 during this period. Goal 3: Validation of MPS platform for monitoring of IAV and MHYOP using field samples. IAV assay was implemented with purified N protein as well as spiked whole virus sample achieving a sensitivity of 128 pM and 103TCID50/ml respectively. MHYOP assays were not run due to the limitation detailed above. Goal 4: Application of MPS platform for detecting SARS-CoV-2. We successfully achieved 5-minute volumetric assay implementation for SARS-CoV-2 (purified) Spike-protein with LOD of 5nM. Surface-based MPS assay was also developed for improving sensitivity results and achieving a LOD of 104pfu/ml (ELISA LOD: 103pfu/ml, Lateral flow assay LOD: 102pfu/ml) with wholevirus spiked serum samples.

Publications

• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Shuang Liang, Denis Tonini, Renata Saha, Jinming Liu, Parsa Yari, Venkatramana D. Krishna, Maxim C-J. Cheeran, Kai Wu, and Jian-Ping Wang. "Static and Dynamic Magnetization Responses of Self-Assembled Magnetic Nanoparticle Chains." The Journal of Physical Chemistry C 127, no. 37 (2023): 18494-18505.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Yari, Parsa, Shuang Liang, Vinit Kumar Chugh, Bahareh Rezaei, Shahriar Mostufa, Venkatramana Divana Krishna, Renata Saha et al. "Nanomaterial-Based Biosensors for SARS-CoV-2 and Future Epidemics." Analytical Chemistry 95, no. 42 (2023): 15419-15449.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: IEM Annual Conference, September 26, 2023. Minneapolis, U.S. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase.
• Type: Conference Papers and Presentations Status: Other Year Published: 2023 Citation: The 68th Annual Conference on Magnetism and Magnetic Materials (MMM 2023), October 30 - November 3, 2023. Dallas, U.S. Title: Systematic Analysis of Parameters Affecting Colorization Performance for Magnetic Particle Spectroscopy (MPS) Applications.
• Type: Book Chapters Status: Published Year Published: 2024 Citation: Vinit Kumar Chugh, Kai Wu, Jian-Ping Wang. Magnetic particle spectroscopy (MPS) biosensor. Book  Magnetic Nanoparticles in Nanomedicine, Elsevier (2024).

Progress 08/01/22 to 07/31/23

Outputs
Target Audience:The direct impact of this project will be on 1) hospitals, clinics, doctors, patients that require fast and on-site COVID-19 diagnosis (Goal 4); 2) and veterinarians and pork producers who require field tests for porcine respiratory diseases (Goals 2&3). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Professional develoment oportunities were provided for two postdocs: Kai Wu, and Venkatraman D Krishna by way of invited talks. Similar opportunities for the two grad students: Vinit Kumar Chucg, and Shuang Liang were also provided through conference attendence and poster presentations. How have the results been disseminated to communities of interest?Apart from the scientific publications, the results were also reported by way of invited talk as mentioned below: Department of Electrical and Computer Engineering, Texas Tech University. Oct 7, 2022. Lubbock, TX, USA.Presenter: Kai Wu. Title:Magnetic Particle Spectroscopy Point-of-Care Device for Disease Diagnosis. Department of Electrical and Computer Engineering, University of Minnesota. Sep 16, 2022. Virtual.Presenter: Kai Wu. Title:Magnetic Biosensors for Point-of-Care (POC) Diagnostics. The National Institute of Standards and Technology (NIST). Apr 20,2023. Virtual. Presenter: Kai Wu. Title:Magnetic Particle Spectroscopy Point-of-Care Device for Disease Diagnosis. What do you plan to do during the next reporting period to accomplish the goals?There are 3 main goals that we will be working towards for the final year of this project: Improved sensitivity implementation by shifting to a surface-bases assay scheme. LOD determination using whole-virus spiked serum samples and real farm sample. These would be needed for validation of technology in real-life settings. Multiplexed detection implementation facilitated by the surface-bases assay implementation.

Impacts
What was accomplished under these goals? Goal 1: Development of a handheld MPS bioassay device with user interface. Two handheld prototypeshave been built with sufficient surface-based assay sensitivity needed for real-world disease diagnostic applications. Sensitivity for volumetric assay implementation still lacks this goal by a sensitivity factor of 40x. User interface (mobile application) has also been developed to enable user communication. Handheld device structure and geometry are also complete. Goal 4: Application of MPS platform for detecting SARS-CoV-2. We successfully achieved 5-minute volumetric assay implementation for SARS-CoV-2 (purified) Spike-protein with LOD of 5nM. Realizing the lower sensitivity implecations of the volumetric assay, a surface-based detection methodology was developed for ananlyte detection.

Publications

• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Arturo di Girolamo, Venkatramana D. Krishna, Kai Wu, Maxim CJ Cheeran, and Jian-Ping Wang. "Frequency and amplitude optimizations for magnetic particle spectroscopy applications." The Journal of Physical Chemistry C 127, no. 1 (2022): 450-460.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Su, Diqing, Kai Wu, Karthik Srinivasan, Zohreh Nemati, Reza Zamani, Vinit Chugh, Renata Saha et al. "Ultra?flexible giant magnetoresistance biosensors for lab?on?a?needle biosensing." Advanced Materials Interfaces 10, no. 7 (2023): 2201417.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Yari, Parsa, Bahareh Rezaei, Clifton Dey, Vinit Kumar Chugh, Naga Venkata Ravi Kumar Veerla, Jian-Ping Wang, and Kai Wu. "Magnetic particle spectroscopy for point-of-care: a review on recent advances." Sensors 23, no. 9 (2023): 4411.
• Type: Journal Articles Status: Published Year Published: 2023 Citation: Chugh, Vinit Kumar, Shuang Liang, Parsa Yari, Kai Wu, and Jian-Ping Wang. "A method for multiplexed and volumetric-based magnetic particle spectroscopy bioassay: mathematical study." Journal of Physics D: Applied Physics 56, no. 31 (2023): 315001.

Progress 08/01/21 to 07/31/22

Outputs
Target Audience:The direct impact of this project will be on 1) hospitals, clinics, doctors, patients that require fast and on-site COVID-19 diagnosis (Goal 4); 2) and veterinarians and pork producers who require field tests for porcine respiratory diseases (Goals 2&3). Efforts: Training Efforts: We have involved and trained three undergrad students, Arturo di Girolamo, Prince Jappah, and Connor Templeton to this project from Spring 2022. This senior design team helped in creation of two handheld devices to be used in our Bioassay applications and they also helped in finalizing the mobile application and PC software development for the handheld devices. Invited Talks: The 13th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, June 14-17, 2022. London, UK. Presenter: Jian-Ping Wang. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Washfree Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. Department of Electrical and Computer Engineering, Texas Tech University. Feb 7, 2022. Lubbock, TX, U.S. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Department of Electrical and Computer Engineering, Texas A&M University. Jan 28, 2022. Virtual. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have involved three senior students, Arturo di Girolamo, Prince Jappah, and Connor Templeton in this project from Spring 2022. This senior design team helped in the creation of two handheld devices to be used in our Bioassay applications and they also helped in finalizing the mobile application and PC software development for the handheld devices. An effort was put in to train the students in using CAD and PCB design tools to further their expertise in the field of Electrical engineering. How have the results been disseminated to communities of interest?Our team's efforts are reported by the following news media: https://cse.umn.edu/ece/feature-stories/alumnus-kai-wu-wins-best-poster-2022-joint-mmmintermag Results are also presented to peers through conferences and publications: The 15th Joint MMM-INTERMAG Conference (2022 Joint), January 10-14, 2022. New Orleans, U.S. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. The 13th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, June 14-17, 2022. London, UK. Presenter: Jian-Ping Wang. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. Department of Electrical and Computer Engineering, Texas Tech University. Feb 7, 2022. Lubbock, TX, U.S. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. Department of Electrical and Computer Engineering, Texas A&M University. Jan 28, 2022. Virtual. Presenter: Kai Wu. Title: Magnetic Biosensors for Point-of-Care (POC) Diagnostics. What do you plan to do during the next reporting period to accomplish the goals?The major goal in the 3rd year of this project will be achieving Goal 2: Application of MPS handheld platform and bench-top system for the detection of IAV and MHYOP, and Goal 3: Validation of MPS platform for monitoring of IAV and MHYOP using field samples. We have made an effort to shift to a Néel relaxation-based sensing mechanism to accommodate the high sensitivity requirements for IAV and MHYOP detection from real field samples. Firstly, the focus would be on the detection of individual analytes to identify the best antibody ratio for bioassay purposes. Once the key ratios have been identified, we will move forward with the multiplexed detection as planned. We are also experimenting with multicore magnetic tracers from different suppliers to help improve the overall detection sensitivity. Our initial experiments suggest further sensitivity improvement by one order (10x).

Impacts
What was accomplished under these goals? During the second year of this award period (8/1/2021 - 7/31/2022), we have accomplished the following goals: Goal 1: Development of a handheld MPS bioassay device with a user interface. We have built two additional handheld MPS devices with improved sensitivity by a factor of 12X from our first generation of prototypes. Corresponding user interface for mobile application has also been updated for user convenience. Goal 2: Application of MPS handheld platform and benchtop system for detection of IAV and MHYOP. The limitation of our present volumetric methodology for multiplexed detection was realized and corresponding steps have been taken by a shift to a Néel relaxation-based detection mechanism to allow us the implementation of multiplexed Biosensing. Goal 4: Application of MPS platform for detecting SARS-CoV-2. We worked on decreasing the assay time through the introduction of heating and agitation to reduce the time required in the MPS bioassay incubation step. This work helped to cut the assay time from several hours to 5 minutes. The corresponding work has been submitted to the journal (unpublished as of 7/31/2022).

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Chugh, V.K., Wu, K., Krishna, V.D., di Girolamo, A., Bloom, R.P., Wang, A. (Y.Q.), Saha, R., Liang, S., Cheeran, M.C., & Wang, J.P. (2021) Magnetic Particle Spectroscopy with One-stage Lock-in Implementation for Magnetic Bioassays with Improved Sensitivities. Journal of Physical Chemistry C, 2021, 125, 31, 1722117231
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Chugh, V.K., Krishna, V.D., di Girolamo, A., Wang, A. (Y.Q.), Saha, R., Liang, S., Cheeran, M.C., & Wang, J.P. (2021) One-step, Wash-free, Nanoparticle Clusteringbased Magnetic Particle Spectroscopy Bioassay Method for Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase. ACS Applied Materials & Interfaces, 2021, 13, 37, 4413644146
• Type: Journal Articles Status: Published Year Published: 2022 Citation: Wu, K., Liu, J., Chugh, V.K., Liang, S., Saha, R., Krishna, V.D., Cheeran, M.C., & Wang, J.P. (2022) Magnetic Nanoparticles and Magnetic Particle Spectroscopy-based Bioassays: A 15-year Recap. Nano Futures, 6, 022001
• Type: Journal Articles Status: Under Review Year Published: 2022 Citation: Wu, K., Chugh, V.K., Krishna, V.D., Wang, Y., Gordon, T.D., Cheeran, M.C., & Wang, J.P. (2022) 5-Minute Magnetic Particle Spectroscopy (MPS)-based Bioassay: Towards Ultra-fast, Convenient, and Large-scale Disease Diagnosis
• Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: The 15th Joint MMM-INTERMAG Conference (2022 Joint), January 10-14, 2022. New Orleans, U.S. Title: Handheld Magnetic Particle Spectroscopy (MPS) for Rapid, One-step, Wash-free Detection of SARS-CoV-2 Spike and Nucleocapsid Proteins in Liquid Phase.

Progress 08/01/20 to 07/31/21

Outputs
Target Audience:Target audiences: The direct impact from this project will be to 1) hospitals, clinics, doctors, patients that requires fast and on-site COVID-19 diagnosis (Goal 4); 2) and veterinarians and pork producers who require field tests for porcine respiratory diseases (Goals 2&3). Efforts: We have involved one high school student, Allison Benjamin, to this project from Summer 2020. She performed qualitative analysis for this team's study assessing the impact of COVID-19 on African healthcare workers. DVM student, Rachel Weidmayer, worked on characterizing the Mycoplasma hypopneumoniae reagents that will be used for the MPS assay. Changes/Problems:Due to the COVID-19 lock down, the mycoplasma and swine work was shut down (Goal 2 & 3). Thus, we continued our work to build a diagnostic assay for SARS-CoV-2 detection (Goal 4). In the first award year, one major challenge that severely delayed our research progress came from the magnetic nanoparticle supplier, who could not provide us with stable and high-performance nanoparticles for MPS-based bioassays. This challenge was overcome with two solutions: 1) instituting a monthly contract with the company to ensure regular supply of magnetic nanoparticles; 2) expanding our vendor pool to search for other sources for magnetic nanoparticle as backup. What opportunities for training and professional development has the project provided?We have involved one high school student, Allison Benjamin, to this project from Summer 2020. She performed qualitative analysis for this team's study assessing the impact of COVID-19 on African healthcare workers The College of Veterinary Medicine Summer Scholars program supported Ms. Rachel Weidmayer, a 3rd year DVM student for the summer (May 2011), to work on characterizing two unique monoclonal antibodies and its ability to detect Mycoplasma hyopneumoniae in clinical samples. She optimized the reagents for a plate ELISA to detect the pathogen in clinical matrix which will be transposed on magnetic nanoparticles for detection with an MPS device How have the results been disseminated to communities of interest?Our team's efforts are reported by several news media: https://vetmed.umn.edu/news/newly-funded-developing-widely-accessible-covid-19-breathalyzer-test https://equimanagement.com/news/umn-vet-school-researchers-working-to-develop-covid-19-breathalyzer-test https://research.umn.edu/inquiry/post/handheld-device-quickly-detect-covid-19 https://spectrum.ieee.org/news-from-around-ieee/biomedical/diagnostics/a-better-faster-cheaper-test-for-covid19 https://kstp.com/coronavirus/university-of-minnesota-researchers-develop-new-portable-coronavirus-testing-device/5707860/ https://cse.umn.edu/college/feature-stories/portable-virus-detection-device-its-way We also built up our team websites to advertise the ongoing work of this project: https://magicoil.ece.umn.edu https://coronavirus.ece.umn.edu Results are also presented to the peers through conferences and publications. What do you plan to do during the next reporting period to accomplish the goals?The major goal in the 2nd year of this project will be achieving the Goal 2: Application of MPS handheld platform and bench-top system for the detection of IAV and MHYOP. We are also enhancing our MPS device designs by upgrading the circuit boards, device structures, to achieve lower the detection limits (increase sensitivity) and optimizing the reagent in the bioassay to enhance analytical sensitivity and specificity.

Impacts
What was accomplished under these goals? During the first year of this award period (8/1/2020 - 7/31/2021), we have accomplished the following goals: Goal 1: Development of a handheld MPS bioassay device with user interface. We have built up 3 prototypes of MPS devices and validated the detection sensitivities of these devices on some basic proteins including SARS-CoV-2 spike and nucleocapsid proteins. The circuit board and structure designs can be found from our publications (ACS Applied Materials & Interfaces,13(7), pp.7966-7976; Frontiers in Biomedical Devices 83549, V001T10A01; arXiv preprint arXiv:2105.12718) The user interface design and animations showing how to use the user application can be found from our publications (ACS Applied Materials & Interfaces,13(7), pp.7966-7976; Frontiers in Biomedical Devices 83549, V001T10A01) as well as YouTube video (https://www.youtube.com/watch?v=E-s5FyblLEM) Goal 4: Application of MPS platform for detecting SARS-CoV-2. We have achieved detection limits of 1.56 nM (equivalent to 125 fmole) and 3.13 nM (equivalent to 250 fmole) for detecting SARS-CoV-2 spike and nucleocapsid proteins, respectively. This work has been submitted to journal (unpublished as of 7/31/2021).

Publications

• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Chugh, V.K., Di Girolamo, A., Liu, J., Saha, R., Su, D., Krishna, V.D., Nair, A., Davies, W., Wang, Y.A. and Cheeran, M.C., 2021. A Portable Magnetic Particle Spectrometer for Future Rapid and Wash-Free Bioassays. ACS Applied Materials & Interfaces, 13(7), pp.7966-7976.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Liu, J., Saha, R., Peng, C., Su, D., Wang, Y.A. and Wang, J.P., 2021. Investigation of Commercial Iron Oxide Nanoparticles: Structural and Magnetic Property Characterization. ACS omega, 6(9), pp.6274-6283.
• Type: Journal Articles Status: Published Year Published: 2021 Citation: Wu, K., Liu, J., Saha, R., Ma, B., Su, D., Chugh, V.K. and Wang, J.P., 2021. Stable and Monodisperse Iron Nitride Nanoparticle Suspension for Magnetic Diagnosis and Treatment: Development of Synthesis and Surface Functionalization Strategies. ACS Applied Nano Materials, 4(5), pp.4409-4418.