Hybridisation mix synthesis in a spiral lab-on-chip device for fast-track microarray genotyping of human pathogens

Johannes R. Peham; Lisa-Maria Recnik; Walter Grienauer; Michael J. Vellekoop; Christa Nöhammer; Herbert Wiesinger-Mayr

doi:10.1117/12.886620

3 May 2011 Hybridisation mix synthesis in a spiral lab-on-chip device for fast-track microarray genotyping of human pathogens

Johannes R. Peham, Lisa-Maria Recnik, Walter Grienauer, Michael J. Vellekoop, Christa Nöhammer, Herbert Wiesinger-Mayr

Author Affiliations +

Proceedings Volume 8068, Bioelectronics, Biomedical, and Bioinspired Systems V; and Nanotechnology V; 806803 (2011) https://doi.org/10.1117/12.886620
Event: SPIE Microtechnologies, 2011, Prague, Czech Republic

Abstract

DNA microarrays can provide bacterial identification, which is crucial for targeted therapy. However they lack rapidness, because of multiple analysis steps. Therefore a fast one-step method for synthesising a hybridisation-ready reagent out of initial bacterial DNA is required. This work presents the combination and acceleration of PCR and fluorescent labelling within a disposable microfluidic chip, fabricated by injection moulding. 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 and a high-yield polymerase leading to a one-step process accelerated by cyclic-flow PCR. Three different bacterial samples (Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) were processed and the bacterial DNA was 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. Furthermore the overall processing time was decreased from 6 hours to 1.5 hours. We showed that a disposable polycarbonate chip, fabricated by injection moulding is suitable for the significant acceleration of DNA microarray assays. The reaction output lead to high-sensitivity bacterial identification in a short time, which is crucial for an early and targeted therapy against infectious diseases.

Citation Download Citation

Johannes R. Peham, Lisa-Maria Recnik, Walter Grienauer, Michael J. Vellekoop, Christa Nöhammer, and Herbert Wiesinger-Mayr "Hybridisation mix synthesis in a spiral lab-on-chip device for fast-track microarray genotyping of human pathogens", Proc. SPIE 8068, Bioelectronics, Biomedical, and Bioinspired Systems V; and Nanotechnology V, 806803 (3 May 2011); https://doi.org/10.1117/12.886620

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available