title: On-chip quantitative PCR using integrated real-time detection by capillary electrophoresis.
abstract: Quantitative PCR (qPCR) has been widely used for the detection and monitoring of a variety of infectious diseases. PCR and CE were integrated into a microfluidic chip that was designed to achieve rapid real-time amplicon sampling, separation, and quantitation without requiring various probes. A novel chip design allows the overlapped execution of PCR and CE, minimizing the time required for CE analysis after each PCR cycle. The performance of the on-chip qPCR method was demonstrated using a 45-minutes model assay protocol for the phiX174 bacteriophage, and the multiplexing capability of the method was demonstrated by adding a second target, E. coli genomic DNA, to the model assay. The results indicate good sensitivity, reproducibility, and linearity over the tested assay range, 50 to 2 × 10(4) copies/25 μL reaction. Based on this performance, the on-chip qPCR method should be applicable to a wide variety of infectious disease detection and monitoring assays with the addition of suitable sample preparation protocols.
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Samuel D
authors.initials: SD
authors.lastname: Chan
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Eto
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Warren
authors.initials: W
authors.lastname: Wu
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Jian Ping
authors.initials: JP
authors.lastname: Zhang
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Ring-Ling
authors.initials: RL
authors.lastname: Chien
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Henry G
authors.initials: HG
authors.lastname: Wada
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Greenstein
authors.affiliation: Wako Life Sciences, Inc, CA, USA.
authors.forename: Shinji
authors.initials: S
authors.lastname: Satomura
mesh_terms: D000465:Algorithms; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D004247:DNA
keywords: CE; MicroTAS; Microfluidic chip; PCR