title: Handheld real-time PCR device.
abstract: 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.
authors.affiliation: KIST-Europe, Microfluidics Group, Campus E7.1, 66111 Saarbrücken, Germany. pavel.neuzil@gmail.com.
authors.forename: Christian D
authors.initials: CD
authors.lastname: Ahrberg
authors.affiliation: National Institute of Standard and Technology (NIST), Center for Nanoscale Science and Technology, 100 Bureau Drive, MS 6201, Gaithersburg, MD 20899-6201, USA.
authors.forename: Bojan Robert
authors.initials: BR
authors.lastname: Ilic
authors.affiliation: KIST-Europe, Microfluidics Group, Campus E7.1, 66111 Saarbrücken, Germany. pavel.neuzil@gmail.com.
authors.forename: Andreas
authors.initials: A
authors.lastname: Manz
authors.affiliation: KIST-Europe, Microfluidics Group, Campus E7.1, 66111 Saarbrücken, Germany. pavel.neuzil@gmail.com and Brno University of Technology (BUT), Central European Institute of Technology (CEITEC), Technická 3058/10, CZ-616 00 Brno, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Neužil
mesh_terms: D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics*; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus