A simple integrated microfluidic device for the multiplexed fluorescence-free detection of Salmonella enterica
Briony C. Strachan,Hillary S. Sloane,Eric Houpt,Jacob C. Lee,Daniel C. Miranian,Jingyi Li,Daniel A. Nelson
Analyst Pub Date : 12/02/2015 00:00:00 , DOI:10.1039/C5AN01969A
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.

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