title: Automated Microfluidic Platform for Serial Polymerase Chain Reaction and High-Resolution Melting Analysis.
abstract: We report the development of an automated genetic analyzer for human sample testing based on microfluidic rapid polymerase chain reaction (PCR) with high-resolution melting analysis (HRMA). The integrated DNA microfluidic cartridge was used on a platform designed with a robotic pipettor system that works by sequentially picking up different test solutions from a 384-well plate, mixing them in the tips, and delivering mixed fluids to the DNA cartridge. A novel image feedback flow control system based on a Canon 5D Mark II digital camera was developed for controlling fluid movement through a complex microfluidic branching network without the use of valves. The same camera was used for measuring the high-resolution melt curve of DNA amplicons that were generated in the microfluidic chip. Owing to fast heating and cooling as well as sensitive temperature measurement in the microfluidic channels, the time frame for PCR and HRMA was dramatically reduced from hours to minutes. Preliminary testing results demonstrated that rapid serial PCR and HRMA are possible while still achieving high data quality that is suitable for human sample testing.
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Weidong
authors.initials: W
authors.lastname: Cao
authors.affiliation: Sobosoft LLC, Catonsville, MD, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Bean
authors.affiliation: Key Tech, Baltimore, MD, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Corey
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Johnathan S
authors.initials: JS
authors.lastname: Coursey
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Kenton C
authors.initials: KC
authors.lastname: Hasson
authors.affiliation: Canon U.S.A., Inc., Melville, NY, USA.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Inoue
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Taisuke
authors.initials: T
authors.lastname: Isano
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Sami
authors.initials: S
authors.lastname: Kanderian
authors.affiliation: Key Tech, Baltimore, MD, USA.
authors.forename: Ben
authors.initials: B
authors.lastname: Lane
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Hongye
authors.initials: H
authors.lastname: Liang
authors.affiliation: Key Tech, Baltimore, MD, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Murphy
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Greg
authors.initials: G
authors.lastname: Owen
authors.affiliation: Canon U.S.A., Inc., Melville, NY, USA.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Shinoda
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA.
authors.forename: Shulin
authors.initials: S
authors.lastname: Zeng
authors.affiliation: Canon U.S. Life Sciences, Inc, Rockville, MD, USA Ivor.Knight@canon.uslifesciences.com.
authors.forename: Ivor T
authors.initials: IT
authors.lastname: Knight
mesh_terms: D057205:Automation, Laboratory / Q000379:methods*; D060005:Genotyping Techniques / Q000191:economics; D006801:Humans; D044085:Microfluidics / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012371:Robotics / Q000379:methods; D013997:Time Factors; D044366:Transition Temperature
keywords: PCR; high-resolution melting analysis; liquid handling; microfluidic