title: Microfluidic Integration of a Cloth-Based Hybridization Array System (CHAS) for Rapid, Colorimetric Detection of Enterohemorrhagic Escherichia coli (EHEC) Using an Articulated, Centrifugal Platform.
abstract: We describe the translation of a cloth-based hybridization array system (CHAS), a colorimetric DNA detection method that is used by food inspection laboratories for colony screening of pathogenic agents, onto a microfluidic chip format. We also introduce an articulated centrifugal platform with a novel fluid manipulation concept based on changes in the orientation of the chip with respect to the centrifugal force field to time the passage of multiple components required for the process. The platform features two movable and motorized carriers that can be reoriented on demand between 0 and 360° during stage rotation. Articulation of the chip can be used to trigger on-the-fly fluid dispensing through independently addressable siphon structures or to relocate solutions against the centrifugal force field, making them newly accessible for downstream transfer. With the microfluidic CHAS, we achieved significant reduction in the size of the cloth substrate as well as the volume of reagents and wash solutions. Both the chip design and the operational protocol were optimized to perform the entire process in a reliable, fully automated fashion. A demonstration with PCR-amplified genomic DNA confirms on-chip detection and identification of Escherichia coli O157:H7 from colony isolates in a colorimetric multiplex assay using rfbO157, fliCH7, vt1, and vt2 genes.
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Matthias
authors.initials: M
authors.lastname: Geissler
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Liviu
authors.initials: L
authors.lastname: Clime
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Xuyen D
authors.initials: XD
authors.lastname: Hoa
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Keith J
authors.initials: KJ
authors.lastname: Morton
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Harold
authors.initials: H
authors.lastname: Hébert
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Lucas
authors.initials: L
authors.lastname: Poncelet
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.
authors.forename: Maxence
authors.initials: M
authors.lastname: Mounier
authors.affiliation: Ontario Laboratory Network, Canadian Food Inspection Agency , Building 22, 960 Carling Avenue, Ottawa, ON K1A 0C6, Canada.
authors.forename: Mylène
authors.initials: M
authors.lastname: Deschênes
authors.affiliation: Ontario Laboratory Network, Canadian Food Inspection Agency , Building 22, 960 Carling Avenue, Ottawa, ON K1A 0C6, Canada.
authors.forename: Martine E
authors.initials: ME
authors.lastname: Gauthier
authors.affiliation: Ontario Laboratory Network, Canadian Food Inspection Agency , Building 22, 960 Carling Avenue, Ottawa, ON K1A 0C6, Canada.
authors.forename: George
authors.initials: G
authors.lastname: Huszczynski
authors.affiliation: Bureau of Microbial Hazards, Health Canada , 251 Sir Frederick G. Banting Driveway, Ottawa, ON K1A 0K9, Canada.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Corneau
authors.affiliation: Ontario Laboratory Network, Canadian Food Inspection Agency , Building 22, 960 Carling Avenue, Ottawa, ON K1A 0C6, Canada.
authors.forename: Burton W
authors.initials: BW
authors.lastname: Blais
authors.affiliation: Life Sciences Division, National Research Council of Canada , 75 de Mortagne Boulevard, Boucherville, QC J4B 6Y4, Canada.|Institut National de la Recherche Scientifique, Centre Énergie, Matériaux, Télécommunications (INRS-EMT) , 1650 Lionel-Boulet Boulevard, Varennes, QC J3X 1S2, Canada.
authors.forename: Teodor
authors.initials: T
authors.lastname: Veres
mesh_terms: D015373:Bacterial Typing Techniques / Q000295:instrumentation; D002498:Centrifugation; D003124:Colorimetry / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D054324:Enterohemorrhagic Escherichia coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D009693:Nucleic Acid Hybridization; D013997:Time Factors
chemical_list: D004269:DNA, Bacterial