Compact optical diagnostic device for isothermal nucleic acids amplification

SENSORS AND ACTUATORS B-CHEMICAL

卷:133

Article

Lee, Szu-Yuan^2,3,4  Huang, Jhen-Gang²  Chuang, Tsung-Liang²  Sheu, Jin-Chuan⁵  Chuang, Yi-Kuang⁶  Holl, Mark^7,8,9  Meldrum, Deirdre R.^7,8,9  Lee, Chun-Nan³  Lin, Chii-Wann^1,2,10,11 

[1] Natl Taiwan Univ, Coll Med, Inst Biomed Engn, Med Microsensor & Syst Lab, Taipei 106, Taiwan

[2] Natl Taiwan Univ, Coll Engn, Taipei 106, Taiwan

[3] Natl Taiwan Univ, Dept Clin Lab Sci & Med Biotechnol, Taipei 106, Taiwan

[4] Mil Taichung Gen Hosp, Dept Lab Examinat, Taichung, Taiwan

[5] Natl Taiwan Univ Hosp, Dept Internal Med, Taipei 100, Taiwan

[6] Natl Taiwan Univ Hosp, Dept Lab Med, Taipei, Taiwan

[7] Arizona State Univ, Ira A Fulton Sch Engn, Phoenix, AZ USA

[8] Arizona State Univ, Dept Elect Engn, Phoenix, AZ USA

[9] Arizona State Univ, Biodesign Inst, Ctr Ecogenom, Phoenix, AZ USA

[10] Natl Taiwan Univ, Inst Elect Engn, Taipei 106, Taiwan

[11] Natl Taiwan Univ, Ctr Nano Sci & Technol, Taipei 106, Taiwan

关键词: LAMP;    isothermal;    diagnostic device;    hepatitis B virus;   

DOI  :  10.1016/j.snb.2008.03.008

来源: Elsevier

【 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. (C) 2008 Elsevier B.V. All rights reserved.

【 授权许可】

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