建立鴿子胚胎成纖維母細胞系用於培養鴿子環狀病毒（PiCV）之研究

Establishment of pigeon embryonic fibroblast cell line for the study of culturing pigeon circovirus （PiCV）

莊肯樺(Chuang Ken-Hua)

指導教授 ： 莊國賓

國立屏東科技大學/獸醫學院/動物疫苗科技研究所/碩士(2021年)

https://doi.org/10.6346/NPUST202100428

摘要

鴿子環狀病毒（Pigeon circovirus, PiCV）為圓環病毒屬，會破壞鴿子的免疫系統，進而使鴿子更容易遭受其他病毒或細菌之感染。由於鴿子常為比賽或表演因素而往返不同地理區域，使得鴿子環狀病毒在全世界快速傳播，影響肉鴿及賽鴿產業，但尚無有效的鴿子環狀病毒疫苗。近期的研究文獻皆顯示，目前沒有開發出培養鴿子環狀病毒的方法，這使得PiCV疫苗的研發更加困難。本研究目的是建立一種利用細胞株培養PiCV的實驗方法，未來可作為研製PiCV疫苗及建立PiCV感染的實驗模型。目前從收集到的病鴿樣本中，發現PiCV在鴿子臟器的感染分布，也第一次在眼睛拭子樣本檢測到PiCV，此實驗結果有助於對PiCV感染途徑的了解。用病鴿組織及PEF（Pigeon embryo fibroblasts）進行病毒培養，最後利用即時定量PCR（Real-time Polymerase chain reaction）檢測病毒核酸量，實驗結果顯示PiCV能在PEF細胞中被培養。將癌化基因送入PEF，使PEF初代細胞永生化，以解決初代細胞有限繼代培養的問題，之後進行多次繼代，實驗結果顯示，轉染癌基因的實驗組細胞狀況比控制組的細胞良好。

關鍵字

鴿子環狀病毒 ； 鴿胚纖維母細胞 ； 鴿子環狀病毒疫苗

並列摘要

The pigeon circovirus （PiCV） belongs to the genus Circovirus. PiCV destroys the immune system of pigeons, that pigeons are more vulnerable to other viruses or bacteria. PiCV infections are spread worldwide due to pigeons often travel to different country for competition or performance. PiCV is a threat to pigeon’s squab and racing pigeons, until now, there is no effective vaccine available against PiCV. According to scientific literatures, there is no available experimental protocol developed to culture PiCV therefore the immunoprophylaxis of this disease is difficult. This study aims to establish an experimental protocol to culture PiCV by using pigeon cell lines, can be used to develop a PiCV vaccine experimental models. At present, we discovery the distribution of PiCV in the pigeon organs from the samples of sick pigeons, and first detected PiCV in the eye swab samples. The results of this experiment are helpful to understand the route of PiCV infection. Got virus from the diseased pigeon and used pigeon embryo fibroblasts （PEF） cells for virus culture. And then used RT-PCR to detect the amount of virus genome. The results show that PiCV can culture in PEF cells. After that, oncogenes have been transfected into PEF cell. The experimental results showed that the cell condition of the experimental group transfected with the oncogene was better than that of the control group.

並列關鍵字

Pigeon circovirus ； Pigeon Embryo Fibroblasts ； PiCV Vaccine

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