Lab-on-a-chip PCR: real time PCR in miniaturized format for HLA diagnostics

Claudia Gaertner; Holger Becker; Nadine Hlawatsch; Richard Klemm; Christian Moche; René Sewart; Rainer Frank; Andreas Willems

doi:10.1117/12.2050233

22 May 2014 Lab-on-a-chip PCR: real time PCR in miniaturized format for HLA diagnostics

Claudia Gaertner, Holger Becker, Nadine Hlawatsch, Richard Klemm, Christian Moche, René Sewart, Rainer Frank, Andreas Willems

Author Affiliations +

Proceedings Volume 9107, Smart Biomedical and Physiological Sensor Technology XI; 91070O (2014) https://doi.org/10.1117/12.2050233
Event: SPIE Sensing Technology + Applications, 2014, Baltimore, MD, United States

Abstract

In case of transplantation or the identification of special metabolic diseases like coeliac disease, HLA typing has to be done fast and reliably with easy-to-handle devices by using limited amount of sample. Against this background a lab-on-a-chip device was realized enabling a fast HLA typing via miniaturized Real-time PCR. Hereby, two main process steps were combined, namely the extraction of DNA from whole blood and the amplification of the target DNA by Real-time PCR giving rise-to a semi-quantitative analysis. For the implementation of both processes on chip, a sample preparation and a real-time module were used. Sample preparation was carried out by using magnetic beads that were stored directly on chip as dry powder, together with all lysis reagents. After purification of the DNA by applying a special buffer regime, the sample DNA was transferred into the PCR module for amplification and detection. Coping with a massively increased surface-to-volume ratio, which results in a higher amount of unspecific binding on the chip surface, special additives needed to be integrated to compensate for this effect. Finally the overall procedure showed a sensitivity comparable to standard Real-time PCR but reduced the duration of analysis to significantly less than one hour. The presented work demonstrates that the combination of lab-on-a-chip PCR with direct optical read-out in a real-time fashion is an extremely promising tool for molecular diagnostics.

Citation Download Citation

Claudia Gaertner, Holger Becker, Nadine Hlawatsch, Richard Klemm, Christian Moche, René Sewart, Rainer Frank, and Andreas Willems "Lab-on-a-chip PCR: real time PCR in miniaturized format for HLA diagnostics", Proc. SPIE 9107, Smart Biomedical and Physiological Sensor Technology XI, 91070O (22 May 2014); https://doi.org/10.1117/12.2050233

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