Miniaturized devices for isothermal DNA amplification addressing DNA diagnostics
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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.
KeywordsMicrofluidic Device Print Circuit Board Computer Numerical Control Temperature Ramp Substrate Thickness
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).
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