- 學位論文
犬疱疹病毒台灣病毒株之分離與gB基因之選殖及定序
Taiwan Isolates of Canine Herpesvirus and Nucleotide Sequence of Glycoprotein B Gene
摘要
在世界上許多國家，都曾經從罹患致死性出血性疾病的初生幼犬，分離出犬疱疹病毒 （ canine herpesvirus；CHV ）。來自不同病毒株的各種基因序列也都陸續被發表。然而，有關台灣分離株的這些資料卻相當缺乏。本研究從自然感染的病例中成功地分離出犬疱疹病毒，並以病理學、病毒學及分子生物學的方法確認。同時將台灣分離株病毒封套醣蛋白 B （ Glycoprotein B；gB ） 的完整基因序列與曾經發表的序列做一個比較。結果各病毒株間gB基因的核苷酸序列顯示高度的相同性百分比，僅有5個單一核酸多型性 （ single nucleotide polymorphism；SNP ），預測轉譯後的胺基酸也僅有3個置換。此外，我們也嘗試對流浪犬氣管樣本進行病毒分離。結果並無法從60個受測樣本中分離出犬疱疹病毒。原位雜交試驗觀察到呈陽性訊號的細胞出現在脾臟的血管腔內，強化了CHV與其他的疱疹病毒感染一樣藉由血行傳播病毒的證據。
並列摘要
Canine herpesvirus ( CHV ) has been isolated from newborn puppies suffering from fatal hemorrhagic disease in many countries all over the world. Sequence data of a variety of CHV genes derived from many strains have been published. However, there is very little information about Taiwan isolates of CHV. In the present study, We have successfully isolated the CHV from natural cases. Pathologic, virologic as well as molecular biologic methods were done to identify the virus. Simultaneously, the complete nucleotide sequences of gB gene were sequenced and compared with those of genes published. There are very high percentage of sequence identities among the strains compared and just only five putative SNPs ( single nucleotide polymorphism ) were found. Three substitutions were detected in the predicted amino acid sequence. In addition, we attempted to isolate virus from the tracheal specimen of stray dogs. CHV could not be isolated from all 60 samples. Cells with positive hybridization signals within the blood vessel of spleen were observed in ISH test conducted. This finding strengthen the evidence that CHV circulate systemically via blood or lymphatic fluid as shown with other herpesviruses.