PCR-free DNA detection using a magnetic bead-supported polymeric transducer and microelectromagnetic traps.
A fluorescent polymeric hybridization transducer supported on magnetic microbeads was investigated for the rapid, ultrasensitive, and sequence-specific detection of DNA. We show that the polymer derivative can be used to detect target DNA directly on magnetic particles by preparing "target-ready" microbeads grafted with the polymer and suitable DNA probes. A detection limit of approximately 200 target copies in a probed volume of 150 muL (1.4 copies/muL) was obtained for a DNA sequence specific to Candida albicans This detection scheme does not require the release of the hybridized target DNA prior to its detection or the labeling or amplification of the nucleic acids. Furthermore, we show that the fluorescence from these biosensing magnetic beads can be read while magnetically confined in a small volume by a microelectromagnetic trap, which offers the possibility of performing both the preconcentration and detection steps simultaneously on the same support. The combination of the fluorescent polymer biosensor with magnetic particle-assisted DNA preconcentration extends the application of this ultrasensitive biosensor to biological samples with complex matrixes and to integrated lab-on-a-chip platforms, where it could be used for fast multitarget DNA detection in point-of-care diagnostics and field analysis.
Dubus
Sébastien
S
Department of Chemistry and Centre d'Optique, Photonique et Laser (COPL), Université Laval, Québec, Canada.
Gravel
Jean-François
JF
Le Drogoff
Boris
B
Nobert
Philippe
P
Veres
Teodor
T
Boudreau
Denis
D
D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics; D015342:DNA Probes; D004271:DNA, Fungal / Q000032:analysis*; D004574:Electromagnetic Fields; D016133:Polymerase Chain Reaction / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D015342:DNA Probes; D004271:DNA, Fungal; D011108:Polymers