Microfluidic cartridges for DNA purification and genotyping processed in standard laboratory instruments

Maximilian Focke; Daniel Mark; Fabian Stumpf; Martina Müller; Günter Roth; Roland Zengerle; Felix von Stetten

doi:10.1117/12.886860

5 May 2011 Microfluidic cartridges for DNA purification and genotyping processed in standard laboratory instruments

Maximilian Focke, Daniel Mark, Fabian Stumpf, Martina Müller, Günter Roth, Roland Zengerle, Felix von Stetten

Proceedings Volume 8066, Smart Sensors, Actuators, and MEMS V; 80660G (2011) https://doi.org/10.1117/12.886860
Event: SPIE Microtechnologies, 2011, Prague, Czech Republic

Abstract

Two microfluidic cartridges intended for upgrading standard laboratory instruments with automated liquid handling capability by use of centrifugal forces are presented. The first microfluidic cartridge enables purification of DNA from human whole blood and is operated in a standard laboratory centrifuge. The second microfluidic catridge enables genotyping of pathogens by geometrically multiplexed real-time PCR. It is operated in a slightly modified off-the-shelf thermal cycler. Both solutions aim at smart and cost-efficient ways to automate work flows in laboratories. The DNA purification cartridge automates all liquid handling steps starting from a lysed blood sample to PCR ready DNA. The cartridge contains two manually crushable glass ampoules with liquid reagents. The DNA yield extracted from a 32 μl blood sample is 192 ± 30 ng which corresponds to 53 ± 8% of a reference extraction. The genotyping cartridge is applied to analyse isolates of the multi-resistant Staphyloccus aureus (MRSA) by real-time PCR. The wells contain pre-stored dry reagents such as primers and probes. Evaluation of the system with 44 genotyping assays showed a 100% specificity and agreement with the reference assays in standard tubes. The lower limit of detection was well below 10 copies of DNA per reaction.

Citation Download Citation

Maximilian Focke, Daniel Mark, Fabian Stumpf, Martina Müller, Günter Roth, Roland Zengerle, and Felix von Stetten "Microfluidic cartridges for DNA purification and genotyping processed in standard laboratory instruments", Proc. SPIE 8066, Smart Sensors, Actuators, and MEMS V, 80660G (5 May 2011); https://doi.org/10.1117/12.886860

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