Fully-integrated microfluidic chips capable of performing DNA amplification from RNA virus, sample transportation, capillary electrophoresis separation,

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Fully-integrated microfluidic chips capable of performing DNA amplification from RNA virus, sample transportation, capillary electrophoresis separation, and on-line optical detection have been developed. The developed system consumes less reagent and sample volumes, and provides higher heating/cooling rates with a more precise temperature control. The proposed microfluidic chip is a powerful tool for the detection of DNA/RNA samples and has significant potential for implementation in micro total analysis systems. A Fully-Integrated Microfluidic Chip for RNA-Virus Detection Fu-Chun Huang 1, Chia-Sheng Liao 2, Gwo-Bin Lee 1,2 1 Department of Engineering Science, 2 Institute of Micro-Electro-Mechanical-System Engineering National Cheng Kung University, Tainan, Taiwan 701 Design and fabrication The study reports a new fully-integrated microfluidic device capable of performing Reverse-Transcription Polymerase Chain Reaction (RT-PCR), transportation of amplified molecular samples, capillary electrophoresis (CE) separation and on-line detection of RNA-based virus. Using three major technologies, including molecular biology, micro-fabrication and micro-fluidics, we develop an innovative micro-total-analysis-system (μ-TAS) for rapid clinical diagnosis. We have successfully demonstrated the detection of the Dengue-2 virus using the hand-held system in a shorter period with less consumption of samples and reagents. The system can perform high heating (20 ℃ /sec) and cooling (10 ℃ /sec) rates. The integrated system has successfully carried out the detection of Dengue-2 virus samples. Abstract Schematic representation of the fully-integrated microfluidic chips for RNA virus detection. (a) Lithography (b) Platinum deposition (c) Gold deposition (d) Insulation layer setup (e) SU-8 mold formation (f) PDMS replication (g) O 2 plasma bonding Simplified fabrication process for the fully- integrated microfluidic chip. RT/PCR reaction chamber PCR reagents RT-PCR reagents Capillary electrophoresis Optical fiber channel Leads of heaters and sensors Peristaltic micro- pneumatic pump RNA RT/PCR reaction chamber sample preparation capillary electrophoresis chip photomultiplier tube computer optical fiber after RT-PCR RT-PCR reagents PCR reagents Photograph of the fully-integrated microfluidic chip after assembly. Experimental (3)(4) (1)(2) (A)(B) (A)Typical PCR thermal cycles generated by the PCR chip. (B)SEM images. (1) A SU-8 mold of two optical fibers and one microchannel, (2) PDMS replication from the SU-8 mold. (3) SU-8 mold of a pneumatic pump, and (4) PDMS replication of the micro- pump. Electrophoregrams of amplified RNA product of Dengue II virus and Hae III digested  x-174 DNA markers. Hg lamp High Voltage Power Supply Optical fiber Computer PMT Focus lens Band-pass filter Temperature control module Transmission line Microfluidic chip Microfluidic control module (A)(B) (A)Schematic representation of the experimental setup for CE detection. Note that excitation and emitted lights are transmitted by buried optic fibers. (B)The PCR and microfluidic control systems. Conclusions Time (min) Fluorescent signal (mV) MEMS design and Micro-fabrication Lab MML 2006