Portable bead-based fluorescence detection system for multiplex nucleic acid testing: a case study with Bacillus anthracis

被引：10

作者：

Gravel, Jean-Francois ^{[1
,2
]}

Geissler, Matthias ^{[3
]}

Chapdelaine, Sebastien ^{[1
,2
]}

Boissinot, Karel ^{[4
]}

Voisin, Benoit ^{[3
]}

Charlebois, Isabelle ^{[4
]}

Poirier-Richard, Hugo-Pierre ^{[1
,2
]}

Gregoire, Alexandre ^{[1
,2
]}

Boissinot, Maurice ^{[4
]}

Bergeron, Michel G. ^{[4
]}

Veres, Teodor ^{[3
]}

Boudreau, Denis ^{[1
,2
]}

机构：

[1] Univ Laval, Dept Chim, Quebec City, PQ G1V 0A6, Canada

[2] Univ Laval, COPL, Quebec City, PQ G1V 0A6, Canada

[3] CNRC, Boucherville, PQ J4B 6Y4, Canada

[4] Univ Laval, Ctr Rech CHUQ, Ctr Rech Infectiol, Quebec City, PQ G1V 4G2, Canada

来源：

MICROFLUIDICS AND NANOFLUIDICS | 2014年 / 16卷 / 06期

关键词：

Biothreat detection; DNA hybridization; Integration; Microfluidic chip; Point-of-care testing; LIGHT-EMITTING-DIODES; NUCLEOTIDE-SEQUENCE; DNA; CHIP; HYBRIDIZATION; ARRAY; PLASMID; LAB; ENCAPSULATION; MUTATIONS;

D O I：

10.1007/s10404-013-1273-y

中图分类号：

TB3 [工程材料学];

学科分类号：

082905 [生物质能源与材料];

摘要：

This paper describes the design, functioning and use of a portable detection platform for multiplex nucleic acid testing. The system features a bead-supported DNA hybridization assay performed inside a microfluidic cartridge. Polystyrene particles modified with DNA capture probes are confined in the detection area and exposed to a solution of fluorescently labeled target DNA strands. The cartridge, fabricated from inexpensive thermoplastic polymers, allows for conducting up to eight assays in parallel. The detection instrument is equipped with a pneumatic module and a manifold lid serving as an interface to mediate fluid displacement on the cartridge. The fluorescence signal deriving from each assay is recorded by a semi-confocal fluorescence reader embedded in the detection platform. The compact design of the instrument and its level of integration make it possible to obtain an analytical result in less than 15 min, while only few manual steps need to be performed in between. A proof-of-concept demonstration involving Cy3-labeled, PCR-amplified genomic DNA confirms the ability to detect Bacillus anthracis in a multiplexed single-assay format using lef and capC genes. Limits of quantification are on the order of 1 x 10(9) copies/mu L for lef targets.

引用

页码：1075 / 1087

页数：13

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