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Disposable microfluidic chip for rapid pathogen identification with DNA microarrays
Authors:Johannes R Peham  Lisa-Maria Recnik  Walter Grienauer  Michael J Vellekoop  Christa Nöhammer  Herbert Wiesinger-Mayr
Affiliation:(1) Health and Environment Department, AIT Austrian Institute of Technology, Muthgasse 11, 1090 Wien, Austria;(2) Aerospace and Advanced Composites GmbH, 2444 Seibersdorf, Austria;(3) Institute of Sensor and Actuator Systems, Vienna University of Technology, Gusshausstrasse 27-29, 1040 Wien, Austria
Abstract:This work presents the combination and acceleration of PCR and fluorescent labelling within a disposable microfluidic chip. The utilised geometry consists of a spiral meander with 40 turns, representing a cyclic-flow PCR system. The used reaction chemistry includes Cy3-conjugated primers leading to a one-step process accelerated by cyclic-flow PCR. DNA of three different bacterial samples (Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) was processed and successfully amplified and labelled with detection limits down to 102 cells per reaction. The specificity of species identification was comparable to the approach of separate PCR and labelling. The overall processing time was decreased from 6 to 1.5 h. We showed that a disposable polycarbonate chip, fabricated by injection moulding is suitable for the significant acceleration of DNA microarray assays. The reaction output led to high-sensitivity bacterial identification in a short time, which is crucial for an early and targeted therapy against infectious diseases.
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