A Rapid and Real-time Nucleic Acid Detection System by Fluorescence Resonance Energy Transfer and Isothermal RNA Amplification

• Main Authors: Jiann-Mou Chen, 陳建謀
• Other Authors: 莊曜宇
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/28118597521477734866

碩士 === 國立臺灣大學 === 電機工程學研究所 === 94 === In this thesis, we develop a detecting method with high accuracy and high sensitivity for specific RNA sequences in isothermal environment. We construct a low-cost and real-time system for detection specific RNA sequences by using this method. Isothermal RNA amplification is a powerful tool to amplify specific sequences of RNA. It can successfully amplify as low as 1 ng of total RNA in 90 minutes. Therefore, we integrate isothermal RNA amplification and Fluorescence Resonance Energy Transfer (FRET) methods to real-time detect fluorescent signals for specific RNA sequences. For increase specificity, two hybridization probes which can bind to the same gene are used in our experiments to increase the accuracy. Base on this study, a novel method with higher accuracy and sensitivity for detecting specific RNA sequence in isothermal environment can be achieved. The results shown that the FRET signal of TRIM28 and UCHL1 genes have been detected in 40-60 min. Therefore, we have successfully setup a new low-cost, real-time detection system, based on combined Isothermal RNA Amplification and FRET.