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Published December 27, 2010 | Published
Journal Article Open

Exploring the limits of ultrafast polymerase chain reaction using liquid for thermal heat exchange: A proof of principle

Abstract

Thermal ramp rate is a major limiting factor in using real-time polymerase chain reaction (PCR) for routine diagnostics. We explored the limits of speed by using liquid for thermal exchange rather than metal as in traditional devices, and by testing different polymerases. In a clinical setting, our system equaled or surpassed state-of-the-art devices for accuracy in amplifying DNA/RNA of avian influenza, cytomegalovirus, and human immunodeficiency virus. Using Thermococcus kodakaraensis polymerase and optimizing both electrical and chemical systems, we obtained an accurate, 35 cycle amplification of an 85-base pair fragment of E. coli O157:H7 Shiga toxin gene in as little as 94.1 s, a significant improvement over a typical 1 h PCR amplification.

Additional Information

© 2010 American Institute of Physics. Received 20 September 2010; accepted 5 November 2010; published online 28 December 2010. The authors gratefully acknowledge financial support from the Boeing Corporation under the SRDMA program and from the NIH under Grant No. R01 HG002644.

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