Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/67406
標題: 應用試管內轉錄及Multiplex Nested RT-PCR檢測多種水生動物法定傳染病病原
Application of in Vitro Transcription and Multiplex Nested RT-PCR to Detect Aquatic Animal Pathogens
作者: 簡茂盛
黃千衿
關鍵字: 應用研究
IHN
漁業類
傳染性造血組織壞死症
病毒性出血性敗血症
多重引子聚合酵素連鎖反應
VHS
Multiplex PCR
摘要: 傳染性造血組織壞死症(Infectious hematopoietic necrosis; IHN)與病毒性出血性敗血症(Viral Hemorrhagic Septicaemia; VHS)為感染多種高經濟性魚類之高致死性與傳染性的病毒性疾病。此二種疾病有許多相似之處,如IHNV及VHSV均被歸屬於Rhabdoviridae。傳染性造血組織壞死病毒,主要是針對鮭鱒魚類之腎臟、脾臟、消化管黏膜下組織及肝臟等造成嚴重之變性及壞死等病變。而病毒性出血性敗血症主要是造成魚類急性全身性出血與組織壞死而導致器官衰竭,二者均被訂為OIE B表列項下水生動物疾病中重要之法定傳染病。被感染之活魚及魚卵為此疾病重要之傳染原,加上近年來國際間活魚及魚卵之貿易頻繁,且病原亦能垂直感染,若沒有快速精確之檢測方法以執行嚴格的檢疫,實易成為疾病傳染的潛在危險因子。而當我國加入WTO後,建立IHNV及VHSV之標準檢測方式,實為刻不容緩之基礎,遂於93年度計畫中,已成功建立多重引子聚合酵素連鎖反應之檢驗方法。然而IHN及VHS病毒乃屬於RNA病毒,其病毒特性本屬不穩定,可疑之病材若因取得或保存不當,加上檢驗過程所經過的RNA粹取,可能導致病毒受到物理性或化學性之傷害,而造成偽陰性,影響第一線檢疫人員之判定。因此新辦計畫中,將採用試管內轉錄之技術,將IHN及VHS陽性質體中的特殊DNA基因片段轉錄為RNA,並利用此RNA作為一內部控制組,來增加診斷作業準確度。並將應用於巢式PCR改進93年度計畫中所研發的multiplex RT-PCR,用以增加其特異性及敏感性。此二種技術的研發,不但可以避免由國外引進活病毒或國內自行培養此二種惡性傳染病病毒之危險性,也節省在台灣尋找陽性病材與向國外索取病原的不便利與時效性,並有助於建立一套快速、準確及簡便的檢疫方法。
Viral hemorrhagic septicemia (VHS) virus and infectious hematopoietic necrosis (IHN) virus are both important infectious agents in farmed rainbow trout, and disease outbreaks can cause major losses in fish farming worldwide. There are many similarities between these two viruses and clinical signs of two diseases. Both VHSV and IHNV are viruses of the Rhabdoviridae family and as the name indicates, VHS shows a hemorrhagic symptom and IHN induces necrosis in hematopoietic organs. However, the pathologic changes observed during IHN infection in salmonids are quite similar to VHS. The organs affected during an infection with both viruses are usually kidney and spleen, though most organs and tissues are affected in the later stages of the disease. Hematopoietic tissue in kidney and white pulp of the spleen are the most frequently affected tissues. Both viral diseases can be transmitted horizontally and vertically with highly morbidity and mortality both in aquaculture fish and in nature. No effective vaccine or any drugs treatment can be applied at this moment. Furthermore, they are on the top list of International Aquatic Animal Health Code" published by the Office International des Epizooties (OIE). Moreover
URI: http://hdl.handle.net/11455/67406
其他識別: 94農科-13.4.1-檢-B1(2)
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=1162699&plan_no=94%E8%BE%B2%E7%A7%91-13.4.1-%E6%AA%A2-B1%282%29&plan_year=94&projkey=PG9410-2430&target=plan&highStr=*&check=0&pnchDesc=%E6%87%89%E7%94%A8%E8%A9%A6%E7%AE%A1%E5%85%A7%E8%BD%89%E9%8C%84%E5%8F%8AMultiplex+Nested+RT-PCR%E6%AA%A2%E6%B8%AC%E5%A4%9A%E7%A8%AE%E6%B0%B4%E7%94%9F%E5%8B%95%E7%89%A9%E6%B3%95%E5%AE%9A%E5%82%B3%E6%9F%93%E7%97%85%E7%97%85%E5%8E%9F
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