神經壞死病毒 (Nervous necrosis virus, NNV) 是造成石斑魚養殖短期間大量致死的疾病，因而造成漁民及經濟重大的損失，研究如何防止及降低養殖戶受神經壞死病毒的傳播是一項重要的工作，瞭解並釐清可能傳播媒介是判定神經壞死病毒傳染途徑研究的重點項目，因此我們收集可能的傳播媒介並以RT–PCR及Real time PCR的方法偵測神經壞死病毒，結果發現鳥糞可能是病毒傳染媒介之一，檢測結果推測鳥類水平傳染的可能性。另一方面同時利用微流體晶片對多病原檢測，可同時檢測三種疾病病原及一宿主基因，以提供高檢測效率並達到快速偵測的效果，以利養殖戶提早做因應。另外探討環境因子對宿主是否影響病毒的致病性，從神經壞死病毒爆發的現象發現，高溫下對魚體爆發神經壞死病毒的情形比低溫嚴重，是否病毒的複製能力與溫度有關，或是宿主本身溫度因子調節病毒的組成有關，因此探討受溫度影響的蛋白，這些蛋白是否與病毒的複製有關，而Hsp90是一個受溫度影響的蛋白，先前石斑的Hsp90 cDNA未被選殖，本篇論文中選殖出石斑Hsp90全長，利用E. coli BL21將載體送入細菌表現，並純化出重組蛋白，生產可專一辨識Hsp90的抗體，探討病毒感染後在各組織中Hsps mRNA表現情形，是否可被用來做評估感染或環境緊迫可能的指標。另一方面探討Hsp90蛋白與病毒複製與組裝的相關性，發現病毒鞘蛋白與Hsp90有結合的現象。

Nervous necrosis virus (NNV) threatens hatchery produciton of larvae and juveniles in Taiwan. Until now, NNV infection causes massive mortality in grouper juveniles, but less was known about its transmission pathway. In this research, we collected the potential transmission pathways and carriers from 21 aquaculture farms near Tainan. We use RT–PCR and Real–time PCR which has sensitivity and specificity for NNV detection to determine the transmission pathway of NNV. We found the horizontal transmission may be through excrement of bird. On the other hand, we also developed a multiplex diagnosis system for NNV by using microfluid chip. This method can detect three pathogens and one host gene in one–step multiplex tube. This rapid detection method can be used to detect and prevent grouper infection at early stage. In recent years, the grouper farming industry has a big problem of pathogen infection, especially at the larva stage of grouper. If we have a good diagnosis method to detect virus or bacteria, we can earlier find and prevent the disease through the pathogenic mechanism and transmission pathway. However, stress response is important during environmental rapid change, especially virus infection or temperature change, pervious research has found Hsp90 as a stress protein, we cloned Hsp90 gene and constructed it to pET–24a and transform to E. coli BL21 to expression recombinant protein for making a antibody specific for Hsp90, Further, we also use RT–PCR to detected tissue of infected and healthy, evaluate various tissue expression and explore virus and Hsps relationship or suitable as a molecular marker. Finally, we find Hsp90 protein interacts with coat protein.

歐明昌。 (2006) ，石斑魚神經壞死病毒B2功能基因分析及檢測方法開發，國立成功大學生物科技研究所碩士論文，台南。

莊蕙菁。 (2005) ，石斑魚神經壞死病毒特性及檢測方法之研究，國立成功大學生物科技研究所碩士論文，台南。

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