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標題: 以大腸桿菌表現水禽環狀病毒外殼蛋白與複製蛋白並應用於血清抗體之調查
Expression Capsid Proteins and Replication Proteins of Waterfowl Circovirus in Escherichia coli and Use in Serological Surveillance
作者: 李洸鉑
Li, Kuang-Po
關鍵字: waterfowl circovirus
Escherichia coli
出版社: 獸醫學系暨研究所
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摘要: 水禽環狀病毒(waterfowl circoviruses)為新興的水禽傳染病原體,病毒易侵犯淋巴組織,造成免疫抑制,增加其它疾病的罹患率。感染的水禽可見發育遲緩、體重減輕、羽毛脫落等臨床症狀,除了依靠臨床症狀診斷,亦可以電子顯微鏡直接觀察病毒顆粒或以聚合酶連鎖反應(polymerase chain reaction, PCR)、dot blot hybridization tests、in situ hybridization、multiply primed rolling circle amplification來偵測病毒核酸。本研究主題為發展抗體檢測技術並應用於抗體的調查,實驗分為三部分,第一部份,以大腸桿菌(E-coli)表現水禽環狀病毒之複製蛋白(replication protein, Rep)與外殼蛋白(capsid protein, Cap),以此二重組蛋白為抗原,建立以multiscreen之western blot檢測水禽環狀病毒抗體。第二部分,應用抗體檢測技術,對台灣地區之水禽養殖場進行水禽環狀病毒血清抗體流行病學之調查,將台灣分成北、中、南、東四個地域進行研究,鵝環狀病毒的抗體陽性率分別為73.9%、70.3%、60%和100%;鴨環狀病毒部分只對東部鴨場進行調查其抗體陽性率為88.8%。第三部分,進行鵝群血清抗體和抗原相互關係的研究,發現可從糞便中檢出病毒核酸,推測水禽環狀病毒可經由糞便污染環境,並且抗體存在的同時,依然可由糞便中檢出病毒,推測抗體無法完全中和病毒。
Waterfowl circovirus is emerging infectious agents of geese and ducks. These viruses could invade lymphoid tissues and lead to immunosuppression increase chance of infecting other diseases. Waterfowls infected exhibited some clinical signs about feather disorder, poor body condition and low weight. Either clinical signs or polymerase chain reaction (PCR), dot blot hybridization, in situ hybridization they can detect the virus DNA, and electron microscope which can directly observe virion could help to diagnose. The study can be divided into three parts; the first part is using E. coli-expression system to produce recombinant replication protein (Rep) and capsid protein (Cap) of waterfowl circovirus to establishing multiscreen western blot system for serological diagnosis. The second part is applying the method in epidemiological study to assay circovirus antibody of waterfowl herds in Taiwan. We divided Taiwan into four regions, the north, the middle, the south and the east, and the positive rate of geese herds is 73.9%, 70.3%, 60% and 100%, respectively. The east positive rate of ducks herds is 88.8%. The third part is that we choose two geese herds to study the relationship between antibodies and viruses. We detected virus DNA in cloacal swab by PCR and we inferred that circovirus can contaminate environment through feces. Besides, we found that antibodies and antigens existed at the same time, it might be that antibodies could not neutralize goose circovirus completely.
其他識別: U0005-1808200612415700
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