碩士 / 國立臺灣大學 / 動物學研究所 / 89 / The goal of this study is to establish a Capture enzyme-linked immunosorbent assay (ELISA) system for the quantification assay of the whitespot syndrome virus (WSSV). This system is designed as that a monoclonal antibody recognizing a major viral envelope protein, VP 28, was used as the primary antibody and the polyclonal antibodies affinity-purified from WSSV-immunized rats sera as the secondary antibody. First, a standard curve was plotted from purified WSSV containing known amounts protein verse the optical density read from capture ELISA. This plot is linear relation between 0.33 and 42.40 ng WSSV/ml hemolymph. And a regression equation of Y= 0.0001X + 0.0332, R2=0.9106 was thus induced. The capture ELISA was subsequently applied to monitor the concentration changes of WSSV proteins in infected shrimp hemolymph.
Three infection trials were proceeded in healthy adult penaeus vannamei using muscle injection, feeding and immersion. The hemolymph was daily collected from individual shrimp and analyzed by the Capture-ELISA system. The concentrations of WSSV proteins within each sample were calculated from the standard curve. Additionally, the cumulative mortality was daily observed from the other replicate of shrimp.
Combining the daily changes of WSSV concentration with the cumulative mortality, a common phenomenon was observed that the peak of WSSV concentration was reached 1 day before the appearance of the highest cumulative mortality amongst the three infection trials. The range of WSSV protein concentration (10.4 to 16 ng WSSV/ml hemolymph) was suggested to be a critical point that could indicate the outbreak of the serve disease and the massive mortality of infected shrimp.
The Capture-ELISA system will be fourthly applied to field cultures. The condition of WSSV infection within culture shrimp populations will be monitored using this system. And a better culture strategy could be adopted by the farmers.
| Identifer | oai:union.ndltd.org:TW/089NTU00312029 |
| Date | January 2001 |
| Creators | Zhao-June Chen, 陳昭君 |
| Contributors | 施秀惠 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 49 |
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