A simple integrated microfluidic device for the multiplexed fluorescence-free detection of Salmonella enterica

Analyst. 2016 Feb 7;141(3):947-55. doi: 10.1039/c5an01969a. Epub 2015 Dec 14.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Lab-On-A-Chip Devices*
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction / instrumentation*
  • Polymethyl Methacrylate / chemistry
  • Pressure
  • Salmonella enterica / genetics
  • Salmonella enterica / isolation & purification*
  • Systems Integration
  • Torque

Substances

  • DNA, Bacterial
  • Polymethyl Methacrylate