Microfluidics is an emerging technology enabling the development of lab-on-a-chip systems for clinical diagnostics, drug discovery and screening, food safety and environmental analysis. Currently, available nucleic acid diagnostic tests take advantage of polymerase chain reaction that allows exponential amplification of portions of nucleic acid sequences that can be used as indicators for the identification of various diseases. At the same time, isothermal methods for DNA amplification are being developed and are preferred for their simplified protocols and the elimination of thermocycling. Here, we present a low-cost and fast DNA amplification device for isothermal helicase dependent amplification implemented in the detection of mutations related to breast cancer as well as the detection of Salmonella pathogens. The device is fabricated by mass production amenable technologies on printed circuit board substrates, where copper facilitates the incorporation of on-chip microheaters, defining the thermal zone necessary for isothermal amplification methods.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price includes VAT (USA)
Tax calculation will be finalised during checkout.
Ahmad F, Hashsham SA (2012) Miniaturized nucleic acid amplification systems for rapid and point-of-care diagnostics: a review. Anal Chim Acta 733:1–15. doi:10.1016/j.aca.2012.04.031
Aracil C, Perdigones F, Moreno JM, Luque A, Quero JM (2015) Portable lab-on-PCB platform for autonomous micromixing. Microelectron Eng 131:13–18. doi:10.1016/j.mee.2014.10.018
Asiello PJ, Baeumner AJ (2011) Miniaturized isothermal nucleic acid amplification, a review. Lab Chip 11:1420–1430. doi:10.1039/c0lc00666a
Erickson D, Li D (2004) Integrated microfluidic devices. Anal Chim Acta 507:11–26. doi:10.1016/j.aca.2003.09.019
Fang X, Liu Y, Kong J, Jiang X (2010) Loop-mediated isothermal amplification integrated on microfluidic chips for point-of-care quantitative detection of pathogens. Anal Chem 82:3002–3006. doi:10.1021/ac1000652
Gill P, Ghaemi A (2008) Nucleic acid isothermal amplification technologies—a review. Nucleosides Nucleotides Nucleic Acids 27:224–243. doi:10.1080/15257770701845204
Haeberle S, Zengerle R (2007) Microfluidic platforms for lab-on-a-chip applications. Lab Chip 7:1094–1110. doi:10.1039/b706364b
Kaprou G et al (2015) Miniaturized devices towards an integrated lab-on-a-chip platform for DNA diagnostics. In: Progress in biomedical optics and imaging, Proceedings of SPIE. doi:10.1117/12.2181953
Kefala IN, Papadopoulos VE, Karpou G, Kokkoris G, Papadakis G, Tserepi A (2015) A labyrinth split and merge micromixer for bioanalytical applications. Microfluid Nanofluid. doi:10.1007/s10404-015-1610-4
Kopp MU, De Mello AJ, Manz A (1998) Chemical amplification: continuous-flow PCR on a chip. Science 280:1046–1048. doi:10.1126/science.280.5366.1046
Lutz S et al (2010) Microfluidic lab-on-a-foil for nucleic acid analysis based on isothermal recombinase polymerase amplification (RPA). Lab Chip 10:887–893. doi:10.1039/b921140c
Mahalanabis M, Do J, Almuayad H, Zhang JY, Klapperich CM (2010) An integrated disposable device for DNA extraction and helicase dependent amplification. Biomed Microdev 12:353–359. doi:10.1007/s10544-009-9391-8
Mahmoudian L, Kaji N, Tokeshi M, Nilsson M, Baba Y (2008) Rolling circle amplification and circle-to-circle amplification of a specific gene integrated with electrophoretic analysis on a single chip. Anal Chem 80:2483–2490. doi:10.1021/ac702289j
Mark D, Haeberle S, Roth G, Von Stetten F, Zengerle R (2010) Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications. Chem Soc Rev 39:1153–1182. doi:10.1039/b820557b
Moschou D et al (2013) Integrated biochip for PCR-based DNA amplification and detection on capacitive biosensors. In: Progress in biomedical optics and imaging—proceedings of SPIE. doi:10.1117/12.2017690
Moschou D, Vourdas N, Kokkoris G, Papadakis G, Parthenios J, Chatzandroulis S, Tserepi A (2014) All-plastic, low-power, disposable, continuous-flow PCR chip with integrated microheaters for rapid DNA amplification. Sens Actuators B Chem 199:470–478. doi:10.1016/j.snb.2014.04.007
Papadakis G, Gizeli E (2014) Screening for mutations in BRCA1 and BRCA2 genes by measuring the acoustic ratio with QCM. Anal Methods 6:363–371. doi:10.1039/c3ay41143e
Papadopoulos VE et al (2014) A passive micromixer for enzymatic digestion of DNA. Microelectron Eng 124:42–46. doi:10.1016/j.mee.2014.04.011
Papadopoulos VE, Kokkoris G, Kefala IN, Tserepi A (2015) Comparison of continuous-flow and static-chamber μPCR devices through a computational study: the potential of flexible polymeric substrates. Microfluid Nanofluid 19:867–882. doi:10.1007/s10404-015-1613-1
Shen K, Chen X, Guo M, Cheng J (2005) A microchip-based PCR device using flexible printed circuit technology. Sens Actuators B Chem 105:251–258. doi:10.1016/j.snb.2004.05.069
Tsougeni K et al (2016) Plasma nanotextured polymeric lab-on-a-chip for highly efficient bacteria capture and lysis. Lab Chip. doi:10.1039/C5LC01217A
Vincent M, Xu Y, Kong H (2004) Helicase-dependent isothermal DNA amplification. EMBO Rep 5:795–800. doi:10.1038/sj.embor.7400200
Vorkas PA, Christopoulos K, Kroupis C, Lianidou ES (2010) Mutation scanning of exon 20 of the BRCA1 gene by high-resolution melting curve analysis. Clin Biochem 43:178–185. doi:10.1016/j.clinbiochem.2009.08.024
Wego A, Richter S, Pagel L (2001) Fluidic microsystems based on printed circuit board technology. J Micromech Microeng 11:528–531. doi:10.1088/0960-1317/11/5/313
Wu A, Wang L, Jensen E, Mathies R, Boser B (2010) Modular integration of electronics and microfluidic systems using flexible printed circuit boards. Lab Chip 10:519–521. doi:10.1039/b922830f
The authors acknowledge Dr. S. Chatzandroulis, NCSR “Demokritos”, for the development of the temperature control unit, to be presented in detail in future work. This work is partly funded by the General Secretariat for Research and Technology/Ministry of Education, Greece and European Regional Development Fund (Sectoral Operational Program: Competitiveness and Entrepreneurship, NSRF 2007-2013)/European Commission (“SYNERGASIA II” project “LambSense: Converging Lamb wave sensors with microtechnologies towards an integrated Lab-on-chip for clinical diagnostics” 11SYN-5-502) and the EC under FP7-ICT-2011.3.2 “LOVE-FOOD: Love wave fully integrated Lab-on-Chip platform for food pathogen detection” (Grant Agreement No: 317742).
About this article
Cite this article
Kaprou, G.D., Papadakis, G., Papageorgiou, D.P. et al. Miniaturized devices for isothermal DNA amplification addressing DNA diagnostics. Microsyst Technol 22, 1529–1534 (2016). https://doi.org/10.1007/s00542-015-2750-x
- Microfluidic Device
- Print Circuit Board
- Computer Numerical Control
- Temperature Ramp
- Substrate Thickness