Author(s)

James M. Trauba, Carl T. Wittwer

Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.

Point-of-care diagnostic testing using PCR requires a device that is fast, economical, and practical. Sub-minute amplification has been demonstrated using high concentrations of primers and polymerase in glass capillaries, but its platform is limited to research use. A system using heated copper blocks to clamp a microfluidic flow-through PCR card fabricated from thin film polycarbonate was modeled, fabricated, and tested. Models show that fluid flowing through a thin-film device clamped between temperature-controlled copper blocks equilibrates to a temperature change in 250 milliseconds. A 2-step, 35 cycle PCR with 1.06 second cycles specifically amplified a 69-base pair fragment from a 450-base pair synthetic DNA template of random sequence with the same performance as the glass capillary system. This system demonstrates the feasibility of <1 minute PCR in an inexpensive, disposable sample container.

PCR, Microfluidics, Diagnostics, Point-of-Care

Trauba, J. and Wittwer, C. (2017) Microfluidic Extreme PCR: <1 Minute DNA Amplification in a Thin Film Disposable. Journal of Biomedical Science and Engineering, 10, 219-231. doi: 10.4236/jbise.2017.105017.