https://scholars.lib.ntu.edu.tw/handle/123456789/83929
標題: | Compact optical diagnostic device for isothermal nucleic acids amplification | 作者: | Lee, Szu-Yuan Huang, Jhen-Gang Chuang, Tsung-Liang JIN-CHUAN SHEU Chuang, Yi-Kuang Holl, Mark Meldrum, Deirdre R. Lee, Chun-Nan CHII-WANN LIN |
關鍵字: | LAMP;isothermal;diagnostic device;hepatitis B virus | 公開日期: | 2008 | 卷: | 133 | 期: | 2 | 起(迄)頁: | 493-501 | 來源出版物: | Sensors and Actuators B: Chemical | 摘要: | We recently reported the successful use of the loop-mediated isothermal amplification (LAMP) reaction for hepatitis B virus (HBV) DNA amplification and its optimal primer design method. In this study, we report the development of an integrated isothermal device for both amplification and detection of targeted HBV DNA. It has two major components, a disposable polymethyl methacrylate (PMMA) micro-reactor and a temperature-regulated optical detection unit (base apparatus) for real- time monitoring of the turbidity changes due to the precipitation of DNA amplification by-product, magnesium pyrophosphate. We have established a correlation curve (R-2 = 0.99) between the concentration of pyrophosphate ions and the level of turbidity by using a simulated chemical reaction to evaluate the characteristics of our device. For the applications of rapid pathogens detection, we also have established a standard curve (R-2 = 0.96 ) by using LAMP reaction with a standard template in our device. Moreover, we also have successfully used the device on seven clinical serum specimens where HBV DNA levels have been confirmed by real-time PCR. The result indicates that different amounts of HBV DNA can be successfully detected by using this device within 1 h. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/128209 http://ntur.lib.ntu.edu.tw/bitstream/246246/128209/1/48.pdf |
DOI: | 10.1016/j.snb.2008.03.008 | SDG/關鍵字: | Acids; DNA; Genes; Health; Light metals; Magnesium; Nucleic acids; Organic acids; Precipitation (chemical); Turbidity; AND detection; DNA amplification; Hepatitis B virus (HBV); Isothermal amplification; Micro reactors; Nucleic acids amplification; Optical detection; Optical diagnostics; Poly methyl methacrylate (PMMA); Precipitation (metallography); Primer design; Pyrophosphate (PPi); Real time monitoring; Turbidity changes; Amplification |
顯示於: | 醫學工程學研究所 |
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