伪狂犬病病毒四川株的分离鉴定及增殖特性

Identification of pseudorabies virus from pigs in Sichuan Province and its proliferation characterization

利用PK-15细胞从四川某猪场分离到1株病毒,经PCR检测、蚀斑纯化、病毒理化试验、血清学试验和动物回归试验证实,该分离株为伪狂犬病病毒(PRV),遂将其命名为PRV-SL;用其分别感染PK-15、Vero、Marc-145、Hela、BHK-21和MDBK细胞,观察病变情况及绘制PRV-SL株在不同动物细胞的一步生长曲线;用TCID50法检测PRV-SL株不同细胞毒的病毒效价。结果显示,PRV-SL株在细胞中均能增殖,但不同细胞之间CPE及病毒效价有所差异:PRV-SL株感染PK-15细胞的病毒效价最高;病毒一步生长曲线显示,PRV-SL株感染BHK-21细胞的隐蔽期最短,在猴源细胞(Vero、Marc-145)上增殖周期最长,增殖周期由短至长依次为BHK-21、MDBK、Hela、PK-15、Vero和Marc-145细胞。结果表明,PRV-SL株具有泛嗜性,在不同动物细胞上的增殖效价和增殖曲线具有明显差异,病毒在不同细胞上的增殖周期长短与病毒效价高低无相关性。

A strain of virus was isolated from the cerebrum samples of the piglets with neurologic symptoms in Sichuan using PK-15 Cells, and the isolated virus was proved to be PRV（named PRV-SL strain） through PCR, plaque purification, physical and chemical properties experiments, serological tests and experimental infection. PRV-SL was used to infect PK-15, Vero, Marc-145, Hela, BHK-21 and MDBK cells respectively,and supernatants were collected at different time points post-infection. One-step growth curves on different animal cells were drawn. Cytopathic effect（CPE） on different cells was observed and TCIDs0 in PK-15 cells was determined. In result, PRV-SL strain could proliferate in different origin cells and cause different CPE. The titer of PRV-SL strain harvested from PK-15 cells was the highest. The onestep growth curve of PRV SL indicated that the BHK-21 cells had the shortest eclipse phase and the monkey origin cells had the longest proliferation period. In conclusion,PRV-SL has the nature of pantropic and shows the different titers and proliferation curves in different animal cells. The length of proliferation peri- od is irrelevant with the level of viral titer.