Here we report one of the smallest real-time polymerase chain reaction (PCR) systems to date with an approximate size of 100 mm × 60 mm × 33 mm. The system is an autonomous unit requiring an external 12 V power supply. Four simultaneous reactions are performed in the form of virtual reaction chambers (VRCs) where a ≈200 nL sample is covered with mineral oil and placed on a glass cover slip. Fast, 40 cycle amplification of an amplicon from the H7N9 gene was used to demonstrate the PCR performance. The standard curve slope was -3.02 ± 0.16 cycles at threshold per decade (mean ± standard deviation) corresponding to an amplification efficiency of 0.91 ± 0.05 per cycle (mean ± standard deviation). The PCR device was capable of detecting a single deoxyribonucleic acid (DNA) copy. These results further suggest that our handheld PCR device may have broad, technologically-relevant applications extending to rapid detection of infectious diseases in small clinics.

KIST Europe Microfluidics Group Campus E7 1 66111 Saarbrücken Germany

KIST Europe Microfluidics Group Campus E7 1 66111 Saarbrücken Germany and Brno University of Technology Technická 3058 10 CZ 616 00 Brno Czech Republic

National Institute of Standard and Technology Center for Nanoscale Science and Technology 100 Bureau Drive MS 6201 Gaithersburg MD 20899 6201 USA

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