Application of gold nanoparticles for the diagnostics of Japanese encephalitis virus.

• Main Authors: Song Yuhan, 宋羽涵
• Other Authors: Huang Suhua
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/86651243832786362311

碩士 === 亞洲大學 === 生物科技學系碩士班 === 100 === Japanese encephalitis virus (JEV) is the pathogen of Japanese encephalitis. It belongs to an enveloped virus of the genus Flavivirus in the family Flaviviridae. Real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) and capture IgM and IgG ELISA are routinely used for the diagnosis of Japanese encephalitis virus (JEV) infection. Gold nanoparticle can increase the sensitivity of in vivo and in vitro diagnosis and is widely used in biomedical and drug treatment development. The aims of this study are the antibody preparation of Japanese encephalitis virus NS1 protein, as well as the application of gold nanoparticle to increase the detection sensitivity of Japanese encephalitis virus NS1 protein. We used gold nanoparticle on two different detection systems including RT-PCR assay and immunochromatography assay. Gold nanoparticles didn’t increased the amplification yield of the PCR products and can’t shortened the PCR reaction time,but gold nanoparticle can increase the sensitivity of immunochromatography assay. We applied anti-JEV NS1 IgG on two parts : one was immobilized in a defined detection zone on a porous nitrocellulose membrane, while the other was conjugated with gold nanoparticles. After mixing biopsy with anti-JEV NS1 IgG-gold nanparticle conjugates, the mixturepassed along the porous membrane by capillary action and bound to the anti-JEV NS1 IgG in the detection zone, in the sandwich format. The test strip then yielded a red color, which is Positive. The sensitivity of the assay for JEV NS1 protein was 10ng/ml. The assays described here were simple, sensitive, and rapid and provided early diagnostic tools for the detection of low level JEV virus in the acute-phase infection.