碩士 / 國立高雄大學 / 生物科技研究所 / 99 / Postlarvae of Macrobrachium rosenbergii infected with white tail disease (WTD) were previously reported in Taiwan. The causative agents have been identified as Macrobrachium rosenbergii nodavirus (MrNV) associated with extra small virus (XSV). In this study, the gene sequence encoding the capsid protein of MrNV and XSV were separately cloned into pGEX-4T-3 expression vector and transformed into Escherichia coli BL-21 strain. After induction, glutathione-S-transferase (GST) tagged MrNV and XSV were obtained with molecular mass of 68 and 43 KDa, respectively. The recombinant proteins were purified by glutathione agarose column before immunization of animal for antibody production. The polyclonal antiserum specific to MrNV and XSV in viral recombinant protein and infected prawn tissue were verified by Western hybridization. By immunodot blot, the detection sensitivities of antibodies were approximately 10ng/ul for recombinant protein GST-MrNV and GST-XSV, respectively. Additional, the MrNV and XSV were detected at a level of dilution 1:2560 and 1:640 in infected tissue, respectively. Additionally, sensitivity of immunodot blot method was less than PCR in this study. The infection of MrNV and XSV in muscle tissue of infected prawn was detected by immunohistochemistry (IHC). The results above revealed that PAbs were specific to MrNV and XSV. The PAbs generated in this study could be used to further develop a simple and rapid immunochromatographic test strip.
| Identifer | oai:union.ndltd.org:TW/099NUK05111009 |
| Date | January 2011 |
| Creators | Chia-yang Chang, 張嘉洋 |
| Contributors | Chun-shun Wang, 王俊順 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 83 |
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