碩士 / 輔仁大學 / 生命科學系碩士班 / 95 / Grouper distribute over the warm water. There are about more than 400 species of grouper around the world. Because of its delicious flavor and rich nutrition value, grouper become a popular commercial fish species. Recently, viruses infection causes tremendous economic losses in aquaculture, but the detection of these viruses is still remain inconvenient. The major infective viruses are iridovirus and nervous necrosis virus, especially Ranavirus, GIV (Grouper iridovirus), SGIV (Singapore grouper iridovirus), Megalocytivirus, OSGIV (Orange-spotted grouper iridovirus and RSIV (Red sea bream iridovirus). The accurate and rapid pathogen detection is very important in the prevention and quarantine of fish disease. Thus, the purpose of this study is to develop sensitive and fast methods of molecular detection for virus diagnosis－loop-mediated isothermal amplification(LAMP) and multiplex RT-PCR. LAMP primer set is designed for GIV major capsid protein (MCP) gene, and the signal can be observed after amplification at 65℃ for 1 hour. The signal can be also observed in the amplification temperature from 57.5 to 70℃. 10-7μg GIV DNA can be detected, the sensitivity is more than 10-fold of traditional PCR result. In multiplex RT-PCR, primers are designed for purine nucleoside phosphorylase (PNP) gene, MCP gene of Ranavirus, GIV and SGIV, ATPase, MCP gene of Megalocytivirus, OSGIV and RSIV, and coat protein (CP) gene of NNV, and it can detect GIV PNP, MCP, NNV CP simultaneously. In order to enhance the sensitivity and accuracy of the method, there should be more clinical samples to do the test. The molecular detection methods of viruses in this study can be used to quarantine all aquaculture fish species and to investigate the infection profile of ocean resources.
| Identifer | oai:union.ndltd.org:TW/095FJU00105015 |
| Date | January 2007 |
| Creators | Wu Hsiu-Ting, 吳琇婷 |
| Contributors | Chang Chi-Yao, 張繼堯 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 64 |
Page generated in 0.0146 seconds