期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 133, 期 2, 页码 493-501出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2008.03.008
关键词
LAMP; isothermal; diagnostic device; hepatitis B virus
资金
- National Science and Technology Program Biotechnology and Pharmaceuticals of National Science Council [NSC 94-2614-B-002-001]
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. (C) 2008 Elsevier B.V. All rights reserved.
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