title: Ultra-rapid flow-through polymerase chain reaction microfluidics using vapor pressure.
abstract: A novel flow-through polymerase chain reaction (PCR) microfluidic system using vapor pressure was developed that can achieve ultra-rapid, small-volume DNA amplification on a chip. The 40-cycle amplification can be completed in as little as 120 s, making this device the fastest PCR system in the world. The chip device is made of a pressure-sensitive polyolefin (PSP) film and cyclo-olefin polymer (COP) substrate which was processed by cutting-work to fabricate the microchannel. The enclosed structure of the microchannel was fabricated solely by weighing the PSP film on the COP substrate, resulting in superior practical application. The vapor pressure in the denaturation zone of the destabilizing flow source was applied to the flow force, and ultra-rapid, efficient amplification was accomplished with a minimal amount of PCR reagents for detection. The flowing rhythm created by vapor pressure minimized the residual PCR products, leading to highly efficient amplification. For field test analysis, airborne dust was collected from a public place and tested for the presence of anthrax. The PCR chip had sufficient sensitivity for anthrax identification. The fastest time from aerosol sampling to detection was theoretically estimated as 8 min.
authors.affiliation: Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, Kagawa, Japan. yu-fuchiwaki@aist.go.jp
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Fuchiwaki
authors.forename: Hidenori
authors.initials: H
authors.lastname: Nagai
authors.forename: Masato
authors.initials: M
authors.lastname: Saito
authors.forename: Eiichi
authors.initials: E
authors.lastname: Tamiya
mesh_terms: D001408:Bacillus anthracis / Q000302:isolation & purification*; D003516:Cycloparaffins / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000096:biosynthesis; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D055548:Vapor Pressure
chemical_list: D003516:Cycloparaffins; D004269:DNA, Bacterial