a fully-integrated microfluidic chip for rna-virus detection
DESCRIPTION
A Fully-Integrated Microfluidic Chip for RNA-Virus Detection. (1). (2). (3). (4). RT-PCR reagents. PCR reagents. after RT-PCR. Computer. Microfluidic chip. High Voltage Power Supply. RT/PCR reaction chamber. capillary electrophoresis chip. sample preparation. PMT. RNA. - PowerPoint PPT PresentationTRANSCRIPT
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 DetectionFu-Chun Huang1, Chia-Sheng Liao2, Gwo-Bin Lee1,2
1Department of Engineering Science, 2Institute of Micro-Electro-Mechanical-System EngineeringNational 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) O2 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
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(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 lensBand-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
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511
72118
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271281
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8721078 1353
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MEMS design and Micro-fabrication LabMML2006