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應用聚合酶連鎖反應檢測水禽中之鵝環狀病毒

Detection of Goose Circovirus in Waterfowls by Polymerase Chain Reaction

摘要


鵝環狀病毒(goose circovirus, GCV)為一新近發現之禽類傳染病原,由於GCV並無任何體外培養系統可供應用,因此無法利用病毒分離來作確診,一般診斷除觀察鶴隻外觀及解剖上之病變外,目前最重要診斷方法為聚合酶連鎖反應(PCR)。本研究依錄所有已發表之GCV核酸序列,設計出一組引子(P1762與P385),並以台灣各地所收集到之水禽臟器檢體進行PCR測試,結果證實引子P1762與P385在病毒檢出率上,優於先前文獻中已發表用於檢測GCV之引子,很適合用於本省GCV之PCR檢測。另外此組引子亦能自正蕃鴨中檢測出環狀病毒之存在,這是首度證實環狀病毒亦能感染鴨隻之報導。核酸序列之比對發現,在正蕃鴨中所發現之環狀病毒有兩種,一種在序列上極類似GCV,其序列相似性為92.7-99.0%之間,另一種則與GCV相距較遠,相似性只有73.5-79.6%。總結而言,本研究所建立之PCR方法,不但可用於GCV之快速檢測,亦能用於檢測鴨隻中之環狀病毒,將來如能利用此技術,對本省之鴨群與鵝群加以廣泛檢測,當可協助瞭解水禽環狀病毒在本省鴨鵝之感染疫情,並有助於相關疾病之預防與治療。

並列摘要


Goose circovirus (GCV) is a newly identified pathogen of geese. Because of the lack of in vitro culture system for GCV, the diagnosis of GCV could not be verified by the method of virus ilation. The observation of clinical symptoms and gross necropsy, as well as the detection of virus by PCR, are important for the diagnosis of this disease. This study designed a set of PCR primers (P1762 and P385) based on the published sequences of GCV, and then tested the efficacy of this set of primers in the detection of GCV collected from different areas of Taiwan. The result of PCR showed that primers P1762 and P385 performed better than all PCR primers published previously; moreover, PCR by using P1762 and P385 could also detect circovirus from Muscovy ducks. This is the first report on the presence of circovirus in ducks. Nucleotide sequences analysis showed that the circovirus found in ducks fell into two groups; the first group had 92.7-99.0% nucleotide sequence identity to GCV whereas the second group had only 73.5-79.6% identity to GCV. In conclusion, the PCR by using primers P1762 and P385 could serve as a rapid method for the detection of circovirus in geese and ducks. The application of this PCR method to the epidemiological study of waterfowl circovirus in Taiwan should contribute to the prevention and treatment of this disease.