Ultrafast real-time PCR with integrated melting curve analysis and duplex capacities using a low-cost polymer lab-on-a-chip system

Rainer Gransee; Tristan Schneider; Deniz Elyorgun; Xenia Strobach; Tobias Schunck; Theresia Gatscha; Christian Winkler; Julian Höth

doi:10.1117/12.2179461

13 May 2015 Ultrafast real-time PCR with integrated melting curve analysis and duplex capacities using a low-cost polymer lab-on-a-chip system

Rainer Gransee, Tristan Schneider, Deniz Elyorgun, Xenia Strobach, Tobias Schunck, Theresia Gatscha, Christian Winkler, Julian Höth

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Proceedings Volume 9487, Smart Biomedical and Physiological Sensor Technology XII; 948706 (2015) https://doi.org/10.1117/12.2179461
Event: SPIE Sensing Technology + Applications, 2015, Baltimore, MD, United States

Abstract

Nucleic amplification using quantitative polymeric chain reaction (qPCR) has become the gold standard of molecular testing. These systems offer both amplification and simultaneous fluorescence detection. An ultrafast microfluidic module (allowing 30 PCR cycles in 6 minutes) based on the oscillating fluid plug concept was previously developed ^[1,2] allowing the amplification of native genomic deoxyribonucleic acid (DNA) molecules. This abstract presents the actual status of the advanced system. The upgraded system generates high quality qPCR amplification plots and additional sensitive melting point analysis comparable to data obtained from commercial real-time cyclers. These features provide the user with all information needed to analyze PCR products. The system uses light emitting diodes (LED) for illumination and a low cost Charge-coupled Device (CCD) camera for optical detection. Image data processing allows the automated process control of the overall system components. The system enables the performance of rapid and robust nucleic acid amplifications together with the integration of real time measurement technology. This allows the amplification and simultaneous quantification of the targeted pathogens. The integration of duplex amplification performance allows the incorporation of the necessary controls into the device to validate the PCR performance. This demonstrator can be run either as fully autonomously working device or as OEM part of a sample-to-answer platform.

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Rainer Gransee, Tristan Schneider, Deniz Elyorgun, Xenia Strobach, Tobias Schunck, Theresia Gatscha, Christian Winkler, and Julian Höth "Ultrafast real-time PCR with integrated melting curve analysis and duplex capacities using a low-cost polymer lab-on-a-chip system", Proc. SPIE 9487, Smart Biomedical and Physiological Sensor Technology XII, 948706 (13 May 2015); https://doi.org/10.1117/12.2179461

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KEYWORDS

Luminescence

Control systems

Light emitting diodes

Microfluidics

Lab on a chip

Ultrafast phenomena

Cameras

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