ASIA unversity:Item 310904400/8637
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    题名: Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus
    作者: 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
    贡献者: Department of Biotechnology
    关键词: Amplification;Computer viruses;Gold;Nanoparticles;Nanostructured materials;Nanostructures;Nucleic acids;RNA;Tissue culture;Viruses;Gold nano particles;Real-time quantitative RT-PCR
    日期: 2008
    上传时间: 2010-04-07 21:18:00 (UTC+8)
    出版者: Asia University
    摘要: 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.
    關聯: Nanotechnology 19:405101
    显示于类别:[生物科技學系] 期刊論文

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