Disposable microfluidic chip for rapid pathogen identification with DNA microarrays

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...

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Veröffentlicht in:Microsystem technologies : sensors, actuators, systems integration actuators, systems integration, 2012-03, Vol.18 (3), p.311-318
Hauptverfasser: Peham, Johannes R., Recnik, Lisa-Maria, Grienauer, Walter, Vellekoop, Michael J., Nöhammer, Christa, Wiesinger-Mayr, Herbert
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Sprache:eng
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Zusammenfassung: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 10 2 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.
ISSN:0946-7076
1432-1858
DOI:10.1007/s00542-011-1401-0