title: Genetic analysis of H1N1 influenza virus from throat swab samples in a microfluidic system for point-of-care diagnostics.
abstract: The ability to obtain sequence-specific genetic information about rare target organisms directly from complex biological samples at the point-of-care would transform many areas of biotechnology. Microfluidics technology offers compelling tools for integrating multiple biochemical processes in a single device, but despite significant progress, only limited examples have shown specific, genetic analysis of clinical samples within the context of a fully integrated, portable platform. Herein we present the Magnetic Integrated Microfluidic Electrochemical Detector (MIMED) that integrates sample preparation and electrochemical sensors in a monolithic disposable device to detect RNA-based virus directly from throat swab samples. By combining immunomagnetic target capture, concentration, and purification, reverse-transcriptase polymerase chain reaction (RT-PCR) and single-stranded DNA (ssDNA) generation in the sample preparation chamber, as well as sequence-specific electrochemical DNA detection in the electrochemical cell, we demonstrate the detection of influenza H1N1 in throat swab samples at loads as low as 10 TCID(50), 4 orders of magnitude below the clinical titer for this virus. Given the availability of affinity reagents for a broad range of pathogens, our system offers a general approach for multitarget diagnostics at the point-of-care.
authors.affiliation: Department of Mechanical Engineering, University of California, Santa Barbara, Santa Barbara, California 93106, USA.
authors.forename: B Scott
authors.initials: BS
authors.lastname: Ferguson
authors.forename: Steven F
authors.initials: SF
authors.lastname: Buchsbaum
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Wu
authors.forename: Kuangwen
authors.initials: K
authors.lastname: Hsieh
authors.forename: Yi
authors.initials: Y
authors.lastname: Xiao
authors.forename: Ren
authors.initials: R
authors.lastname: Sun
authors.forename: H Tom
authors.initials: HT
authors.lastname: Soh
mesh_terms: D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification; D004563:Electrochemistry / Q000295:instrumentation*; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000302:isolation & purification*; D057230:Limit of Detection; D008280:Magnetics; D046210:Microfluidic Analytical Techniques; D010614:Pharynx / Q000821:virology*; D019095:Point-of-Care Systems; D011489:Protein Denaturation; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000737:chemistry; D013048:Specimen Handling
chemical_list: D004277:DNA, Single-Stranded; D012367:RNA, Viral; D012261:Ribonucleoproteins