Compact optical diagnostic device for isothermal nucleic acids amplification

Szu Yuan Lee, Jhen Gang Huang, Tsung Liang Chuang, Jin Chuan Sheu, Yi Kuang Chuang, Mark Holl, Deirdre Meldrum, Chun Nan Lee, Chii Wann Lin

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

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 (R2 = 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 (R2 = 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.

Original languageEnglish (US)
Pages (from-to)493-501
Number of pages9
JournalSensors and Actuators, B: Chemical
Volume133
Issue number2
DOIs
StatePublished - Aug 12 2008

Keywords

  • Diagnostic device
  • Hepatitis B virus
  • Isothermal
  • LAMP

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

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  • Cite this

    Lee, S. Y., Huang, J. G., Chuang, T. L., Sheu, J. C., Chuang, Y. K., Holl, M., Meldrum, D., Lee, C. N., & Lin, C. W. (2008). Compact optical diagnostic device for isothermal nucleic acids amplification. Sensors and Actuators, B: Chemical, 133(2), 493-501. https://doi.org/10.1016/j.snb.2008.03.008