标题: 应用奈米金粒子之于猪瘟病毒E2抗原之检测
Application of the gold nanoparticle in the detection of the swine fever virus E2 antigen
作者: 伍登洋
Wu, Teng-Yang
袁俊杰
Yuan, Chiun-Jye
生物科技学系
关键字: 猪瘟;E2 抗原;奈米金;酵素免疫;快速检测试片;swine fever;E2 antigen;gold nanoparticles;Enzyme-linked immunosorbent assay;rapid test strip
公开日期: 2010
摘要: 典型猪瘟(CSF)是一种猪瘟病毒所引起的高度致命的猪传染性疾病,导致许多国家养猪业的大量经济损失。因此,如何开发简单、快速、有效诊断典型猪瘟病毒的分析方法极为重要,以减少养猪业所受的损害与冲击。当猪只被感染后,会大量产生抗体对抗猪瘟病毒的E2醣蛋白,因而E2醣蛋白被视为诊断典型猪瘟的关键生物标的。因此,我们开发了一系列利用抗体(Ab)、山葵过氧化酶 (HRP)与奈米金粒子键结作为侦测平台(Ab/HRP-AuNPs)的方法,例如酵素连结免疫吸收试验(ELISA)、快速检测试片、电流式免疫感测器来检测猪瘟病毒E2抗原。在直接型ELISA和三明治型ELISA实验中,Ab/HRP-AuNPs对于E2抗原展现良好的灵敏度。直接型ELISA的线性侦测范围从1.95±0.10到31.25±2.05 fmoles, 而三明治型ELISA 则是从125±13.30到4000±583.43 fmoles。至于侦测极限,直接型ELISA和三明治型ELISA分别是1.95±0.10及61.25±5.36 fmoles。关于免疫快速诊断试片的开发,对于E2抗原的侦测范围从2.25到22 pmoles,侦测极限达2.25 pmoles。
Classical swine fever (CSF) is a highly contagious and fatal disease of swine due to the infection of CSF virus (CSFV), causing considerable economic loss in swine industry of many countries. Therefore, the development of the simple, quick and effective analytic methods for the detection of CSFV infection is essential in reducing the damage of swine industry. The antibody specifically against glycoprotein E2 of CSFV (E2 antigen) was found to be largely booted in the infected pigs. Accordingly, E2 antigen was identified to be the key biomarker for the diagnosis of CSF. Therefore, various detection methods for the detection of E2 antigen based on the antibody and horseradish peroxidase (HRP) conjugated gold nanoparticles (Ab/HRP-AuNPs), including ELISA and rapid test strip, were design, developed and characterized in this study. The Ab/HRP-AuNPs exhibited a high sensitivity toward E2 antigen on the direct or sandwich type ELISA assay format. The linear detection range is 1.95±0.10 to 31.25±2.05 fmoles with direct ELISA and 125±13.30 to 4000±583.43 fmoles with sandwich ELISA. The detection limit was 1.95±0.10 fmoles and 62.5±5.36 fmoles for direct and sandwich ELISA, respectively. The flow assay strip was developed for a quick detection of E2 antigen exhibiting a detection range from 2.25 to 22 pmoles with a detection limit of 2.25 pmoles.
URI: http://140.113.39.130/cdrfb3/record/nctu/#GT079828513
http://hdl.handle.net/11536/47721
显示于类别:Thesis