Microfluidic device architecture for electrochemical patterning and detection of multiple DNA sequences.
Electrochemical biosensors pose an attractive solution for point-of-care diagnostics because they require minimal instrumentation and they are scalable and readily integrated with microelectronics. The integration of electrochemical biosensors with microscale devices has, however, proven to be challenging due to significant incompatibilities among biomolecular stability, operation conditions of electrochemical sensors, and microfabrication techniques. Toward a solution to this problem, we have demonstrated here an electrochemical array architecture that supports the following processes in situ, within a self-enclosed microfluidic device: (a) electrode cleaning and preparation, (b) electrochemical addressing, patterning, and immobilization of sensing biomolecules at selected sensor pixels, (c) sequence-specific electrochemical detection from multiple pixels, and (d) regeneration of the sensing pixels. The architecture we have developed is general, and it should be applicable to a wide range of biosensing schemes that utilize gold-thiol self-assembled monolayer chemistry. As a proof-of-principle, we demonstrate the detection and differentiation of polymerase chain reaction (PCR) amplicons diagnostic of human (H1N1) and avian (H5N1) influenza.
Pavlovic
Elizabeth
E
Department of Materials, Chemistry and Biochemistry, University of California, Santa Barbara, California 93111, USA.
Lai
Rebecca Y
RY
Wu
Ting Ting
TT
Ferguson
Brian S
BS
Sun
Ren
R
Plaxco
Kevin W
KW
Soh
H T
HT
D001483:Base Sequence; D015374:Biosensing Techniques; D004247:DNA / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods
D015342:DNA Probes; D004279:DNA, Viral; D004247:DNA