Analysis of Bluetongue Virus mRNA in Baby Hamster Kidney Cells and Sheep Tissues by Real-time Reverse Transcription Polymerase Chain Reaction

以即時反轉錄聚合酶連鎖反應檢測幼齡倉鼠細胞及綿羊組織內的藍舌病病毒傳訊RNA

李璠(Fan Lee) ； 羅婷(Ting Lo) ； 王汎熒(Fun-In Wang)

《臺灣獸醫學雜誌》 37卷2期 (2011/06) Pp. 83-92

https://doi.org/10.7009/TVJ.201106.0083

摘要

藍舌病病毒屬於里奧病毒科的環形病毒屬，為藍舌病的病原。本研究係利用即時反轉錄聚合酶連鎖反應量測藍舌病毒基因轉錄傳訊RNA量，探討臺灣分離的BTV2/KM/2003病毒株在幼齡倉鼠細胞株與及在經福馬林固定的威染綿羊組織內基因轉錄的特性。結果顯示在感染的細胞株中，轉錄量最高的藍舌病毒基因依序為NS2、VP4、VP7。不同的是，在感染綿羊組織內轉錄量最高的為3個非結構蛋白基因NS1、NS2、NS3。在所有受檢組識中，NS1、NS2、NS3基因在舌、唇、耳翼及小腦的表現量略高於其他組織，與之前的實驗結果相呼應，進一步顯示本土分離株BTV2/KM/2003對柯麗黛綿羊為低毒力。因為舌、唇、耳翼及小腦皆集中於頭臉部，顯示接種病毒的組織穿透力較差，因此離接種部位（對側耳翼）較為接近的組織其NS1-3基因的表現量優於較遠端的器官。另一個可能因素即是BTV2對上述頭臉組織有較高的親和性。

關鍵字

藍舌病病毒 ； 基因轉錄 ； 即時反轉錄連鎖反應

並列摘要

Bluetongue virus (BTV), a member of the genus Orbivirus of the family Reoviridae, is the causative agent of bluetongue. The present study further characterizes the replication of Taiwan prototype BTV2/KM/2003 in baby hamster kidney cell line BHK-21 and in formalin-fixed paraffin-embedded tissues of an infected sheep. Messenger RNA transcribed from the BTV genes was measured by a real-time reverse transcription polymerase chain reaction. In the infected BHK-21 cells, the three most transcribed genes were NS2, VP4, and VP7 genes. The abundance of BTV mRNA species in the tissues of infected sheep demonstrated a different pattern, wherein NS1-3 genes were most frequently transcribed. Among all tissues examined, NS1-3 genes were detected at relatively higher levels, indicated by the lower Ct values, in the tongue, haired lip, ear pinna and cerebrum, consistent with the low virulence of the Taiwan BTV2/KM/2003 to Corriedal sheep as previously characterized. The tongue, haired lip, ear, and cerebrum are located primarily in the craniofacial regions and are closer to the inoculation site of contralateral ear pinna. The inoculated BTV should have low or moderate penetration capability, so that the viral gene expressions in the more distant tissues were relatively low, despite the establishment of viremia and the permissiveness of tissues to the virus. Alternatively BTV may have higher tropism for these craniofacial tissues.

並列關鍵字

bluetongue virus ； gene transcription ； real-time reverse transcription polymerase chain reaction