(348d) Affordable Rapid PCR-Based Biodetection at the Point of Care Via Rayleigh-Bénard Convection

Nucleic acid testing approaches like the PCR and its variants are analytical gold standards that play a critical role in diagnosing and monitoring infectious diseases. But PCR-based testing is primarily performed in laboratory settings because the required thermocycling instrumentation is neither amenable to nor validated for operation in a portable format, limiting rapid detection in remote locations and low-middle income countries. A lack of quantitative data to support validation as accepted laboratory methods is a barrier to adopting portable systems for routine use at the point of care (POC). Here we describe a microfluidic POC PCR platform that overcomes this barrier by exploiting the ability to isothermally perform PCR via natural convection [1-2]. This platform can be easily and cost-effectively manufactured without complex equipment and processes, dramatically increasing simplicity, portability, and affordability. The condition to achieve 100% repeatability is identified, providing critical quantitative data to support the adoption of portable convective thermocycling platforms as a centerpiece of validated laboratory methods. We evaluate the platform’s performance by determining the detection limit, the minimum reaction time required, and the potential for multiplex analysis. We apply this platform to demonstrate rapid replication of targets associated with multiple pathogens, including SARS-CoV-2. These results are comparable to or surpass those obtained using the conventional benchtop PCR thermocyclers, supporting the high potential of this platform to enable affordable POC nucleic acid-based analysis.

[1] R. Muddu et al., “Chaotically accelerated polymerase chain reaction by microscale Rayleigh-Bénard convection.” Angewandte Chemie, International ed.) Vol. 50 (2011): 3048-52.

[2] A. Priye et al., “Microscale chaotic advection enables robust convective DNA replication.” Analytical Chemistry Vol. 85 (2013) 10536-41.