Short communication: Localization of Morbillivirus Nucleic Acid in a Pygmy Sperm Whale (Kogia breviceps) by In Situ-Reverse Transcriptase-Polymerase Chain Reaction

要本研究結合聚合酶鏈鎖反應的敏感性與原位雜交之細胞定位功能，以原位反轉錄聚合酶鏈鎖反應進行一隻擱淺小抹香鯨肺臟組織中之麻疹病毒核酸定位。目標基因為麻疹病毒之磷蛋白基因，並以igoxigenin-11-dUTP進行增幅。可清楚地於肺泡上皮細胞、以及肺泡間隔、細支氣管腔與肺泡腔之單核炎症細胞內被發現麻疹病特異性核酸之陽性訊號。與前人以免疫組織化學染色研究結果相較，原位反轉錄聚合酶鏈鎖反應於此病例提供較多診斷病理學上之資訊。此研究亦表明藉由原位反轉錄聚合酶鏈鎖反應之高敏感性與不易造成樣本污染之特性，該方法可用於麻疹病毒感染之疾病發生學研究。

關鍵字

原位反轉錄聚合酶鏈鎖反應；麻疹病毒；小抹香鯨

並列摘要

Combining the sensitivity of PCR and cell-localizing ability of in situ hybridization, the in situ RT-PCR technique was applied to localize cetacean morbillivirus nucleic acid in the lung of a stranded pygmy sperm whale (Kogia breviceps). Primers targeted to phosphoprotein gene of morbillivirus and digoxigenin-11-dUTP was incorporated into the amplicons. Positive signals indicating the presence of morbilliviral specific nucleic acid was clearly found in the type I pneumocyte and mononuclear inflammatory cells in the alveolar septa, space of bronchioli and alveoli. In comparison to previous immunohistochemical findings, in situ RT-PCR provided more direct evidence of causative agents in this case. The result also suggests that in situ RT-PCR with its strength of high sensitivity and natural resistance to sample contamination could be a useful method in the pathogenesis investigation of the morbillivirus infections.

並列關鍵字

in situ RT-PCR ； morbillivirus ； pygmy sperm whale

國際替代計量

Short communication: Localization of Morbillivirus Nucleic Acid in a Pygmy Sperm Whale (Kogia breviceps) by In Situ-Reverse Transcriptase-Polymerase Chain Reaction

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