Compact optical diagnostic device for isothermal nucleic acids amplification
Resource
Sensors and Actuators B: Chemical v.133 n.2 pp. 493-501
Journal
Sensors and Actuators B: Chemical
Journal Volume
133
Journal Issue
2
Pages
493-501
Date Issued
2008
Date
2008
Author(s)
Lee, Szu-Yuan
Huang, Jhen-Gang
Chuang, Tsung-Liang
Chuang, Yi-Kuang
Holl, Mark
Meldrum, Deirdre R.
Lee, Chun-Nan
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 (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.
Subjects
LAMP
isothermal
diagnostic device
hepatitis B virus
SDGs
Other Subjects
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
Type
journal article
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