title: A simple integrated microfluidic device for the multiplexed fluorescence-free detection of Salmonella enterica.
abstract: Rapid, inexpensive and simplistic nucleic acid testing (NAT) is pivotal in delivering biotechnology solutions at the point-of-care (POC). We present a poly(methylmethacrylate) (PMMA) microdevice where on-board infrared-mediated PCR amplification is seamlessly integrated with a particle-based, visual DNA detection for specific detection of bacterial targets in less than 35 minutes. Fluidic control is achieved using a capillary burst valve laser-ablated in a novel manner to confine the PCR reagents to a chamber during thermal cycling, and a manual torque-actuated pressure system to mobilize the fluid from the PCR chamber to the detection reservoir containing oligonucleotide-adducted magnetic particles. Interaction of amplified products specific to the target organism with the beads in a rotating magnetic field allows for near instantaneous (<30 s) detection based on hybridization-induced aggregation (HIA) of the particles and simple optical analysis. The integration of PCR with this rapid, sequence-specific DNA detection method on a single microdevice presents the possibility of creating POC NAT systems that are low cost, easy-to-use, and involve minimal external hardware.
authors.affiliation: Dept of Chemistry, McCormick Road, University of Virginia, Charlottesville, VA 22904, USA. landers@virginia.edu jl8pr@virginia.edu.
authors.forename: Briony C
authors.initials: BC
authors.lastname: Strachan
authors.forename: Hillary S
authors.initials: HS
authors.lastname: Sloane
authors.forename: Eric
authors.initials: E
authors.lastname: Houpt
authors.forename: Jacob C
authors.initials: JC
authors.lastname: Lee
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Miranian
authors.forename: Jingyi
authors.initials: J
authors.lastname: Li
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Nelson
authors.forename: James P
authors.initials: JP
authors.lastname: Landers
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D056656:Lab-On-A-Chip Devices; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D019904:Polymethyl Methacrylate / Q000737:chemistry; D011312:Pressure; D019779:Salmonella enterica / Q000235:genetics / Q000302:isolation & purification*; D018511:Systems Integration; D019415:Torque
chemical_list: D004269:DNA, Bacterial; D019904:Polymethyl Methacrylate