Microfluidic Integration of a Cloth-Based Hybridization Array System (CHAS) for Rapid, Colorimetric Detection of Enterohemorrhagic Escherichia coli (EHEC) Using an Articulated, Centrifugal Platform

被引:25
作者
Geissler, Matthias [1 ]
Clime, Liviu [1 ]
Hoa, Xuyen D. [1 ]
Morton, Keith J. [1 ]
Hebert, Harold [1 ]
Poncelet, Lucas [1 ]
Mounier, Maxence [1 ]
Deschenes, Mylene [2 ]
Gauthier, Martine E. [2 ]
Huszczynski, George [2 ]
Corneau, Nathalie [3 ]
Blais, Burton W. [2 ]
Veres, Teodor [1 ,4 ]
机构
[1] Natl Res Council Canada, Div Life Sci, Boucherville, PQ J4B 6Y4, Canada
[2] Canadian Food Inspect Agcy, Ontario Lab Network, Ottawa, ON K1A 0C6, Canada
[3] Hlth Canada, Bur Microbial Hazards, Ottawa, ON K1A 0K9, Canada
[4] Inst Natl Rech Sci, Ctr Energie, Mat, Telecommun INRS EMT, Varennes, PQ J3X 1S2, Canada
关键词
LAB; EXTRACTION; ASSAYS;
D O I
10.1021/acs.analchem.5b03085
中图分类号
O65 [分析化学];
学科分类号
070302 [分析化学];
摘要
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 degrees 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.
引用
收藏
页码:10565 / 10572
页数:8
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