Increase the ability to perform nucleic acid analysis in a variety of PON settings
The project objectives include:
The development of single-use PCR-based nucleic acid testing will catalyze a radical shift in biomanufacturing and diagnostics by enabling gold standard nucleic acid analysis to be routinely performed in virtually any PON setting.
In addition, the completion of this project will lay the groundwork for efforts to organize manufacturing partners, initiate regulatory processes, and engage in customer discovery so that deployment can begin immediately upon project completion.
This project decentralizes high-fidelity molecular testing by integrating a battery-powered, convective thermocycling instrument with co-packaged reagents, reducing diagnostic hardware capital expense by 70–85% compared to stationary lab equipment. This portable platform targets a 50% reduction in amplification time, enabling real-time nucleic acid analysis at the point of need for vaccine manufacturing and clinical diagnostics. By simplifying the workflow, the system is projected to lower operational user error and "test-failure" rates by 20–30%, drastically cutting the cost-per-actionable-result in non-traditional settings.
The team applied microscale thermal convection to enable isothermal accelerated PCR, overcoming a critical barrier to portable PCR-based testing
The team discovered how DNA amplification can be chemically programmed by manipulating the interplay between the PCR bio-chemistry and the microscale convective flow field
The team discovered that chemical programmability is counterintuitively enabled by elevated amplicon GC contents that are significantly higher than the established recommended levels
The team discovered that chemical programmability enables accelerated isothermal DNA amplification by prolonging extension, the rate-limiting step in the replication process, independent of all other stages in a temperature cycle, making it possible to achieve 100% repeatability at speeds that rival ultra-fast PCR instruments while using lower reagent concentrations than conventional protocols
The team achieved rapid, highly repeatable PCR and RT-PCR with high portability potential, paving the way to deploy lab-quality nucleic acid-based diagnostics in decentralized settings
The team produced prototype PCR tubes incorporating the newly discovered optimal design parameters and performed preliminary studies incorporate real-time melting curve analysis
Kim, M., Ravisankar, V., Hassan, Y. A., & Ugaz, V. M. (2024). Biochemically Programmable Isothermal PCR. Advanced Science, 11(41). https://doi.org/10.1002/advs.202404688
Hazeri, A.H., Marsool, W., Ravisankar, V., Contreras-Naranjo, J.C., and Ugaz, V.M. Quantitative Analysis of DNA Helicity State within a Convective PCR Chamber through CFD Simulations. Poster 174ap. 2024 Annual Meeting of the American Institute of Chemical Engineers, San Diego, CA, October 28, 2024
Kim, M.G, Ravisankar, V., Hassan, Y.A., and Ugaz, V.M. Isothermal Microfluidic PCR Enables Accelerated Pathogen Detection and Diagnostics. Poster M216.j. The 28th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2024. Montreal, Canada, October 14, 2024
Marsool, W., Hazeri, A.H., Ramamirtham, S.A.M., Ravisankar, V., Contreras-Naranjo, J.C., and Ugaz, V.M. Towards Rapid Real-Time DNA Analysis Using Quantitative Fluorescence in Convective PCR. Poster 174ao. 2024 Annual Meeting of the American Institute of Chemical Engineers, San Diego, CA, October 28, 2024
Ugaz, V., Presenter, ARP-23 Rapid Single-use PCR-based Nucleic Acid Testing, NIIMBL National Meeting, Washington, D.C., July 28, 2022.
Ugaz, V.M. Chemically Programmable Isothermal PCR. Poster PS2-12. 2024 NIIMBL National Meeting, June 26, 2024
Kim, M.G., Ravisankar, V., Kim, M.G., Hassan, Y.A., and Ugaz, V.M., Presenter, Accelerating Pathogen Detection: Towards Portable, Rapid Isothermal PCR Diagnostics. Paper 436b. 2024 Annual Meeting of the American Institute of Chemical Engineers, San Diego, CA, October 29, 2024
Ugaz, V.M. Invited Keynote, Chemically Programmable Isothermal PCR Enables Rapid Nucleic Acid-based Bioanalysis at the Point of Need. Materials & Tools for Developing POC & Rapid Dx 2023. Laguna Hills, CA, November 30, 2023
Ugaz, V.M. Invited Oral Presentation, Chemically Programmable Isothermal PCR. NIIMBL Portfolio Snapshot of Member-Led Projects (Part II). 2024 NIIMBL National Meeting, June 26, 2024.
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Texas A&M University System
