Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/93082
標題: Host specificity and construction of a live attenuated vaccine strain of Muscovy duck parvovirus
正番鴨小病毒宿主特異性研究與減毒活毒疫苗株之構築
作者: 顏廷穎
Ting-Ying Yen
關鍵字: 正番鴨小病毒
疫苗
Muscovy duck parvovirus
vaccine
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摘要: Waterfowl parvovirus infection is a highly contagious and fatal disease of goslings and ducklings. According to host specificity and viral nucleotide sequences, waterfowl parvoviruses are divided into two groups: the goose parvovirus (GPV) group and the Muscovy duck parvovirus (MDPV) group. Two major outbreaks of waterfowl parvovirus infection occurred in Taiwan in the last three decades. The first was caused by GPV whereas the second by MDPV. In Taiwan, a live attenuated vaccine strain is available for the control of GPV, but no vaccine strain is available for MDPV. The lack of vaccine strain hampers the control of MDPV. Infectious clone methodology is a valuable tool to study the pathogenic mechanisms of parvoviruses, but no infectious clone of MDPV is yet available. In this study, a plasmid clone containing the full-length genome of MDPV was constructed using the TA cloning methodology. This MDPV clone was found to be infectious after transfection of primary Muscovy duck embryo fibroblast cells and passage in embryonated Muscovy duck eggs. Site-directed mutagenesis showed that the K75N mutation in the VP1 protein of MDPV resulted in the partial attenuation of the virus. Previous study shows that GPV causes the disease in both geese and Muscovy ducks whereas MDPV causes the disease only in ducks but not in geese. In this study, the white Roman geese were experimentally inoculated with MDPV. PCR analysis showed that the geese inoculated with MDPV shed virus from cloacalafter inoculation. Western blot analysis showed that the geese inoculated with MDPV produced antibodies against MDPV. Taken together, these results indicated that the white Roman goose is a host for infection and viral shedding of MDPV. In summary, this studydevelops a simple method for constructing infectious clone of MDPV and this method can facilitate development of vaccines against diseases caused by MDPV. Further, this study also demonstrates that MDPV infection occurred not only in ducks but also in geese. These results are important for the control of waterfowl parvovirus infection in the field.
水禽小病毒感染症是一個高傳染力與高致病力的疾病,主要好發於雛鴨及雛鵝,根據其感染宿主不同及基因序列之差異,水禽小病毒可分為鵝小病毒及正番鴨小病毒,在台灣過去三十年曾經爆發二次水禽小病毒大流行,一次為鵝源的小病毒所造成;另一次是鴨源的小病毒所造成。台灣地區目前已有鵝小病毒疫苗可供使用,但仍無正番鴨小病毒疫苗可使用,造成後者在防疫上之困難。 infectious clone對於探討小病毒的致病機轉是相當有力的工具,但過去研究中仍然未有正番鴨小病毒infectious clone被建構出,在本研究中,我們利用 TA cloning的方法得到完整的正番鴨小病毒基因體序列,並且證實,如將正番鴨小病毒infectious clone轉染正番鴨胚蛋初代細胞,再接種鴨胚蛋,可以產出具有傳染力的正番鴨小病毒。另一方面,也經由點突變的實驗發現,將病毒VP1蛋白的第75個胺基酸進行點突變後(Lysine變成Asparagine),能得到具有部分減毒效果的病毒株,可用於研發正番鴨小病毒活毒疫苗。 過去研究顯示鵝小病毒所引起疾病,主要是發生雛鵝及正番鴨的疾病,而正番鴨小病毒所引起之疾病則是發生於鴨而不發生於鵝。本研究將白羅曼鵝接種正番鴨小病毒進行感染實驗,並接種病毒後不同時間點收集雛鵝糞便檢體進行PCR分析,以檢測是否有感染及排毒現象,本研究也利用西方墨點轉漬法進行觀察感染正番鴨小病毒的白羅曼鵝是否有抗體,將結果交叉比對後,證實白羅曼鵝確實可以被正番鴨小病毒感染及有排毒的現象產生。 本研究結果不僅提供一個研發正番鴨小病毒疫苗之快速方法,而且可以釐清正番鴨小病毒感染宿主特異性。此外,infectious clone技術,可促進對於正番鴨小病毒致病機制的了解,這些研究結果對於未來在控制正番鴨小病毒的田間感染具有主要參考價值。
URI: http://hdl.handle.net/11455/93082
文章公開時間: 10000-01-01
Appears in Collections:微生物暨公共衛生學研究所

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