Microfluidic Extreme PCR: <1 Minute DNA Amplification in a Thin Film Disposable

• 作者: Carl T. Wittwer James M. Trauba
• 来源期刊: 生物医学工程(英文)
• 学科: 医学
• 关键词: PCR MICROFLUIDICS DIAGNOSTICS POINT-OF-CARE
• 年，卷（期）: 2017,（5）
• 页码范围: 219-231
• 页数: 13页
• 分类号: R73

摘要

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.