魚病的控制為養殖中重要的一環，尤其是病毒性之疾病往往造成養殖業之重大損失，近年來本地魚類以腦神經壞死病毒 (nervous necrosis virus)引起的危害最大。台灣石斑養殖產業發展也因此受到很大的阻礙，近年來也有相關研究指出虹彩病毒可能為另一重要的病原。由於虹彩病毒感染致死的魚隻外表並不會有明顯的病徵，在診斷上不容易辨別。過去針對虹彩病毒所設計的特異性引子使用上仍有改進空間，故本研究的第一部份，為改進虹彩病毒的分子診斷的方法。major capsid protein (MCP)為虹彩病毒外鞘蛋白的主要成份，其基因在不同品系的病毒間差異不大，適合用來作為發展universe primer以作為診斷依據，我們以MCP基因為基礎建立以 nested-PCR 診斷方法可有效的用來檢測受到虹彩病毒感染的魚隻，藉由本法可使病毒之偵測極限可達到兩個copy。本研究的第二部份則為發展虹彩病毒與腦神經壞死病毒雙價次單位口服疫苗，在虹彩病毒方面選用MCP當作開發次單位疫苗的標的物，在腦神經壞死病毒則為外鞘蛋白，希望利用生物包埋口服疫苗的平台系統將表現抗原的表現載體可使幼魚同時接受兩種主要疾病的疫苗接種，達到防治疾病的目的。

In recent years, several viral infected diseases have caused a heavy loss in aquaculture industry in Taiwan. One high economic value marine fish, groupers, was also effected by disease problem and the extreme problems of diseases was caused by nervous necrosis virus and iridovirus. The most serious problem of grouper is nervous necrosis virus NNV. But some reports was shown that iridovirus might also a potential pathogen in grouper. But it is hard to be detected due to the small amount of the virus gene in sample. In the first part of the thesis, we design specific primers by viral major capsid protein (MCP) gene and detect viral existence. It could be successful identify even 2 copies of viral DNA by nested PCR. In the secondary part of this research, we try to develop an effective bivalent subunit vaccine and overcome viral caused diseases in groupers. Through bioencapsulated oral vaccine system, we try to elevate fish basic immune response to defend virus infection as well as increased the larvae survival rate in groupers. Subsequence, we try to modulated the optimal ratio between two subunit proteins and assessed humoral response by ELISA in order to expand a effective oral subunit vaccine.

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