title: A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection.
abstract: Solid-phase PCR (SP-PCR) has become increasingly popular for molecular diagnosis and there have been a few attempts to incorporate SP-PCR into lab-on-a-chip (LOC) devices. However, their applicability for on-line diagnosis is hindered by the lack of sensitive and portable on-chip optical detection technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples.
authors.affiliation: Laboratory of Applied Micro and Nanotechnology (LAMINATE) National Food Institute, Technical University of Denmark (DTU-Food), Denmark; Institut Charles Gerhardt UMR 5253 CNRS-UM, Université de Montpellier, Montpellier 34000, France.
authors.forename: Tran Quang
authors.initials: TQ
authors.lastname: Hung
authors.affiliation: Laboratory of Applied Micro and Nanotechnology (LAMINATE) National Food Institute, Technical University of Denmark (DTU-Food), Denmark.
authors.forename: Wai Hoe
authors.initials: WH
authors.lastname: Chin
authors.affiliation: BioLabChip group, Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, DK-2800 Kgs. Lyngby, Denmark. Electronic address: sun.yi@nanotech.dtu.dk.
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.affiliation: BioLabChip group, Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, DK-2800 Kgs. Lyngby, Denmark.
authors.forename: Anders
authors.initials: A
authors.lastname: Wolff
authors.affiliation: Laboratory of Applied Micro and Nanotechnology (LAMINATE) National Food Institute, Technical University of Denmark (DTU-Food), Denmark.
authors.forename: Dang Duong
authors.initials: DD
authors.lastname: Bang
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004867:Equipment Design; D005453:Fluorescence; D005504:Food Analysis / Q000295:instrumentation; D006801:Humans; D061925:Immobilized Nucleic Acids / Q000737:chemistry / Q000235:genetics; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D012480:Salmonella Infections / Q000175:diagnosis / Q000382:microbiology*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D061925:Immobilized Nucleic Acids
keywords: High sensitivity; Lab-on-a-chip; Microlens array; Solid phase PCR; Supercritical angle fluorescence