title: A low-cost, label-free DNA detection method in lab-on-chip format based on electrohydrodynamic instabilities, with application to long-range PCR.
abstract: In order to evolve from a "chip in the lab" to a "lab on a chip" paradigm, there is still a strong demand for low-cost, portable detection technologies, notably for analytes at low concentrations. Here we report a new label-free DNA detection method with direct electronic read, and apply it to long-range PCR. This method uses a nonlinear electrohydrodynamic phenomenon: when subjected to high electric fields (typically above 100 V cm(-1)), suspensions of large polyelectrolytes, such as long DNA molecules, create "giant" dynamic concentration fluctuations. These fluctuations are associated with large conductivity inhomogeneities, and we use here a contact-mode local conductivity detector to detect these fluctuations. In order to decouple the detection electronics from the high voltage excitation one, an original "doubly symmetric" floating mode battery-operated detection scheme was developed. A wavelet analysis was then applied, to unravel from the chaotic character of the electohydrodynamic instabilities a scalar signal robustly reflecting the amplification of DNA. As a first proof of concept, we measured the products of the off-chip amplification of 10 kbp DNA from lambda phage DNA, achieving a sensitivity better than 100 fg DNA in the original 50 μl sample. This corresponds to the amplification products of less than 100 initial copies of target DNA. The companion enabling technologies developed to implement this new concept, i.e. the doubly symmetric contact conductivity detection and wavelet analysis, may also find various other applications in lab-on-chips.
authors.affiliation: UMR 168 Institut Curie, Centre National de la Recherche Scientifique et Université Pierre et Marie Curie, 26 Rue d'Ulm, 75005, Paris, France.
authors.forename: Mohamed Lemine Youba
authors.initials: ML
authors.lastname: Diakité
authors.forename: Jerôme
authors.initials: J
authors.lastname: Champ
authors.forename: Stephanie
authors.initials: S
authors.lastname: Descroix
authors.forename: Laurent
authors.initials: L
authors.lastname: Malaquin
authors.forename: François
authors.initials: F
authors.lastname: Amblard
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Viovy
mesh_terms: D010582:Bacteriophage lambda; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D004560:Electricity; D057446:Hydrodynamics; D056656:Lab-On-A-Chip Devices / Q000191:economics*; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000295:instrumentation*; D058067:Wavelet Analysis
chemical_list: D004279:DNA, Viral; D004247:DNA