摘要
在构建了伪狂犬病病毒上海株的缺失载体 pgEI-GFP 基础上,将 pgEI-GFP 转染感染了 PRV-SH 株的 BHK-21细胞,待出现 80%以上的细胞病变时收获病毒,并以绿色荧光蛋白为标志,通过蚀斑法得到纯化重组病毒gE-/gI-/GFP+ 缺失株。研究了该缺失株的的安全性、在细胞上的生长特性以及对断奶仔猪的安全性、免疫原性等生物学特性。试验结果显示,缺失了 gE-和 gI-后,不影响其在 RK 细胞上的生长状况和病毒的滴度。该疫苗株对小鼠的半数致死量比亲本毒低且对家兔的致死性的时间延长了,这表明该疫苗的毒力比亲本毒有所下降。该缺失株对断奶仔猪安全,无不良接种反应,接种断奶仔猪能抵御高剂量 PRV-SH 株强毒的感染,攻毒后试验猪的发热期、散毒天数均低于对照组。该缺失株接种仔猪后在试验期间一直维持较高水平的中和抗体。
On the basis of construction the transfer vector pgEI-GFP of Pseudorabies virus SH strain BHK-21, which was infected with PRV-SH for 1-2h, were tansfected with the complex of pgEI-GFP and DOTAPA deletion mutant was selected and purified 3-4 times in BHK-21 cell through GFP. In the research here, we investigated the gE-/gI-/GFP+ PRV vaccine strain growth properties in cultured cells, its safety for rabbits, its LD50 for mice, its safety and immunity for postweaning piglets, and its biological properties. The results suggested that gE and gI genes deletion may not affect PRV’s propagation in cultured cells, nor the typical PRV plaque forming. The investigated results demonstrated that the virulence of gE-/gI-/GFP+ was reduced when compared with that of PRV-SH. The inoculation of gE-/gI-/GFP+ couldn’t harm the post-weaning pigs, couldn’t induce the viral spread in surroundings. The period of fever and the number of days of existing virus in treated group pigs were less than that in the control group after big dose of RPV-SH were used to equally attack the treated and control group pigs through ear veins 5 weeks later. gE-/gI-/GFP+could always induce the high titer PRV neutralizing antibodies throughout the experiment.
出处
《中国病毒学》
CSCD
2004年第6期607-611,共5页
Virologica Sinica
基金
上海市农委重点攻关课题(98-05-7)
