TITLE:
Microfluidic Extreme PCR:
AUTHORS:
James M. Trauba, Carl T. Wittwer
KEYWORDS:
PCR, Microfluidics, Diagnostics, Point-of-Care
JOURNAL NAME:
Journal of Biomedical Science and Engineering,
Vol.10 No.5,
May
18,
2017
ABSTRACT: 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