| Summary: | 碩士 === 國立交通大學 === 生物科技學系 === 99 === 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.
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