高病原性雞傳染性華氏囊病病毒之分離與定性

Isolation and Characterization of Very Virulent Infectious BursalDisease Viruses in Taiwan

蔡向榮(1Hsiang. Jung Tsai) ； 林地發(Dih-Fa Lin) ； 呂榮修(Yong-Siu Lu) ； 蔡睦宗(Mo-Chung Tsai)

《中華民國獸醫學會雜誌》 24卷1期 (1998/03) Pp. 42-51

https://doi.org/10.7009/JTCSVS.199803.0042

摘要

台灣在1992年雞傳染性華氏囊病（IBD）大流行時及其後於台灣各地區臨床病例所分離之病毒株共10株，以免疫擴散法、電子顯微鏡檢查法、反轉錄聚合酶鏈反應（RT-PCR）、及抗原捕捉式酵素結合免疫分析法（ELISA）等証明為IBD病毒。分離毒株以核酸探針pOH405及pOH632証明為第一血清型IBD病毒；以血清型單特異性抗血清中和試驗，亦証明分離毒株屬於第一血清型IBD病毒，並與歐洲分離之高病原性IBD（VVIBD）毒株有相似之抗原性。分離毒株接種於4及8週齡SPF雞，可引起典型之IBD症狀及剖檢與組織病變，對8週齡SPF雞可引起40-50%的死亡率，對4週齡SPF雞可引起50-100%的死亡率，另外對4週齡肉雞可引起6 -20%的死亡率。分離毒株中僅有一株在盲目繼代3及5代後，可分別馴化於雞胚胎纖維芽細胞及vera株化細胞上。台灣分離株及1株日本分離之VVIBD毒株之RT-PCR產物，以限制酶Sspl及Taql切割有相同之切割圖譜。綜合上述結果證明台灣自1992年以來所流行之IBD毒株為VVIBD毒株，這些毒株不論在病原性、抗原性、核酸構造、及細胞培養特性上皆與歐日所報告之VVIBD毒株相似。

關鍵字

高病原性雞傳染性華氏囊病病毒 ； 反轉錄聚合酶鏈反應 ； 限制酶切割圖譜

並列摘要

Ten infectious bursal disease viruses (IBDV) were isolated in Taiwan during an epizootic of IBD with high mortality in 1992 and since. IBDVs were isolated by SPF chickens or chicken embryos, and were confirmed by agar gel precipitin test, a commercial antigen capture ELISA kit, and a polymerase chain reaction. The viruses were proved to be serotype 1 IBDV by hybridization with nucleic acid probe pOH632 but not with serotype 2 specific probe pOH405. They also showed to have similar antigenicity with European very virulent IBDV (VVIBDV) by having a similar neutralization index with a serotype 1 monospecific antiserum. The IBD isolates induced 50-100% and 40-50% mortality in 4-week-old and 8-week-old SPF chickens, respectively. They also caused a 6-20% mortality in broilers at 4 weeks of age. Out of ten IBDV isolates only one isolate can be adapted in primary chicken embryo fibroblast and vero cells after 3-5th blind passages. The amplified cDNA products 01 six Taiwan VVIBDV isolates and a reference Japanese VVIBDV isolate were digested by either Ssp I or Taq I restriction enzyme. The results suggest the prevailing IBDV in Taiwan may had a similar nucleotide sequence with European and Japan VVIBDV strains.

並列關鍵字

Very virulent infectious bursal disease ； VVIBO ； Virulence ； Polymerase chain reaction ； Restrictionenzyme