病毒性神經壞死症(Viral nervous necrosis disease,VNN disease)為近年來臺灣石斑魚養殖上極為嚴重的疾病，其致病原為神經壞死病毒(Nervous Necrosis Virus,NNV)，此病毒感染會造成石斑魚苗及幼魚大量死亡，死亡率往往高達90%~100%，造成臺灣石斑養殖業極大的損失。
神經壞死病毒屬於野田病毒科(Nodaviridae)中的betanodavirus屬，此科還有另一屬為alphanodavirus，主要感染植物和昆蟲。以往對神經壞死病毒其蛋白質的研究多偏向於鞘蛋白，但從alphanodavirus的研究可知在病毒感染的過程中，subgenomic RNA3所轉譯出的非結構蛋白質B2對病毒複製扮有重要功能，其與對抗寄主的防禦系統以利病毒複製有關。在此研究中，探索了解石斑魚神經壞死病毒的B2的特性，發現其在細胞內的表現位置為細胞質和細胞核皆存在；而在B2功能分析方面，則發現其具有與雙股RNA結合的能力，並可抑制寄主細胞內的RNAi機制，而更進一步利用點突變的B2蛋白作分析，發現B2蛋白質中第53個位置的arginine與其結合雙股RNA的能力有重要關係。
另外，本實驗室與成大工科所合作開發神經壞死病毒微流體檢測晶片，期望能利用為流體晶片的優點來改善檢驗方法。此研究中使用直接溶解RT-PCR以節省時間和人力；而用RNA1的3端做為檢測標的可更有效早期偵測到病毒的感染，搭配上微流體晶片的高靈敏度，將能達到使檢測方法更快速有效的訴求。

In recent years, viral nervous necrosis disease （VNN）is the most acute disease of hatchery-reared grouper in Taiwan . The causative agent of VNN was nervous necrosis virus （NNV）and the mortality of grouper larvae and juvenile during disease outbreak were usually above 90％.
NNV belongs to the beta subgroup of the virus family nodavirade. Nodavirade has another subgroup alpha-nodavirus which infect inscets and plants. From the study of alpha-nodavirus, it is known that the non-structural B2 protein plays an important role in viral replication and B2 can inhibit the RNA interference system of the host cell. However, the function of beta-nodavirus B2 protein is not clear. In the study, I found that the localization of B2 is in the cytoplasm and nucleus,and B2 can binds to dsRNA and inhibits RNAi of the host cell. Advanced study by dsRNA binding assay of B2 with point mutation, I also found that the fifty-third arginine of B2 protein is involved in the dsRNA binding activity.

Additionally, for effective prevention of nervous nercrosis disease, establishing a fast and accurate diagnosis method is very important work. Microfluidic chip has been characterized by convenient and quick operation. Therefore, our laboratory try to develop a diagnosis method on microfluidic chip. In this study, we use direct-lysis RT-PCR and take RNA1 3’ as detected target for the goal of fast and effective diagnosis.

莊蕙菁(2005)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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