Handheld real-time PCR device

Here we report one of the smallest real-time polymerase chain reaction (PCR) systems to date with an approximate size of 100 mm x 60 mm x 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 approximate to 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.