An integrated microfluidic chip for DNA/RNA amplification, electrophoresis separation and on-line optical detection
An integrated microfluidic chip for DNA/RNA amplification, electrophoresis separation and on-line optical detection
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DOI：
10.1002/elps.200600458
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发表时间：
2006-08-01
期刊：
影响因子：
2.9
通讯作者：
Lee, Gwo-Bin
中科院分区：
文献类型：
--
作者：
Huang, Fu-Chun;Liao, Chia-Sheng;Lee, Gwo-Bin
本研究提出了一种集成微流控芯片，能够自动进行DNA/RNA（脱氧核糖核酸/核糖核酸）扩增、电动进样分离和核酸产物在线光学检测。在该装置中，DNA/RNA样品首先使用基于微机器的PCR模块或反转录PCR （RT-PCR）模块进行复制，然后通过气动微泵输送到样品储存库。样品随后被电动驱动到微通道中，在那里它们被电泳分离，然后通过埋置的光纤进行光学检测。集成微流控芯片的各种模块由廉价的生物相容性材料制成，如PDMS，聚甲基丙烯酸甲酯和钠石灰玻璃。所提出的设备的功能通过其成功应用于基于dna的肺炎链球菌细菌检测和基于rna的登革热-2病毒检测来证明。结果表明，PCR/RT-PCR模块的低热惯性降低了样品和试剂的消耗，缩短了反应时间。由于人工干预较少，后续的DNA分离和检测可以在自动模式下进行。本研究提出的集成微流控装置对分子生物学、遗传分析、传染病检测和其他生物医学应用领域做出了重要贡献。
This study presents an integrated microfluidic chip capable of performing DNA/RNA (deoxyribonucleic acid/ribonucleic acid) amplification, electrokinetic sample injection and separation, and on-line optical detection of nucleic acid products in an automatic mode. In the proposed device, DNA/RNA samples are first replicated using a micro-machine-based PCR module or reverse transcription PCR (RT-PCR) module and then transported by a pneumatic micropump to a sample reservoir. The samples are subsequently driven electrokinetically into a microchannel, where they are separated electrophoretically and then detected optically by a buried optical fiber. The various modules of the integrated microfluidic chip are fabricated from cheap bio-compatible materials, such as PDMS, polymethylmethacrylate, and soda-lime glass. The functionality of the proposed device is demonstrated through its successful application to the DNA-based bacterial detection of Streptococcus pneumoniae and the RNA-based detection of Dengue-2 virus. It is shown that the low thermal inertia of the PCR/RT-PCR modules reduces the sample and reagent consumption and shortens the reaction time. With less human intervention, the subsequent DNA separation and detection could be performed in an automatic mode. The integrated microfluidic device proposed in this study represents a crucial contribution to the fields of molecular biology, genetic analysis, infectious disease detection, and other biomedical applications.