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    Please use this identifier to cite or link to this item: http://asiair.asia.edu.tw/ir/handle/310904400/8637


    Title: Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus
    Authors: Huang, Su-Hua;Yang, Tsuey-Ching;Tsai, Ming-Hong;Tsai, I-Shou;Lu, Huang-Chih;Chuang, Pei-Hsin;Wan, Lei;Lin, Ying-Ju;Lai, Chih-Ho;Lin, Cheng-Wen
    Contributors: Department of Biotechnology
    Keywords: Amplification;Computer viruses;Gold;Nanoparticles;Nanostructured materials;Nanostructures;Nucleic acids;RNA;Tissue culture;Viruses;Gold nano particles;Real-time quantitative RT-PCR
    Date: 2008
    Issue Date: 2010-04-07 21:18:00 (UTC+8)
    Publisher: Asia University
    Abstract: Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples.
    Relation: Nanotechnology 19:405101
    Appears in Collections:[生物科技學系] 期刊論文

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