猪繁殖与呼吸综合征病毒培养工艺的优化

Optimization of culture process for porcine reproductive and respiratory syndrome virus

[目的]优化猪繁殖与呼吸综合征病毒(CH-1a株)在高度敏感的Marc-145克隆细胞上的产毒工艺,完全实现病毒的无血清培养工艺并增加病毒的收次,以提升疫苗的产能和品质。[方法]根据猪繁殖与呼吸综合征病毒(CH-1a株)在Marc-145克隆细胞上的产毒特性,优化维持液中的不同血清浓度、不同pH值以及接毒收获工艺,以提高病毒滴度、降低病毒液中的血清蛋白含量以及增加病毒的收次。[结果]维持液pH控制在7.8~8.2时,采用全换液方式进行无血清培养,一收与二收的混合液病毒滴度达8.13lgTCID_(50)/mL,病毒载量达1.73×10^(7)copies/mL。[结论]建立猪繁殖与呼吸综合征病毒(CH-1a株)在Marc-145克隆细胞上最佳产毒工艺并实现无血清培养,为提升猪繁殖与呼吸综合征病毒(CH-1a株)灭活疫苗的产能和品质提供参考。

[Objective]To optimize the production process of porcine reproductive and respiratory syndrome virus(CH-1a strain)on highly sensitive Marc-145 clone cells,so as to realize the virus serum-free culture and increase the collection of virus,in order to increase the production and quality of vaccines.[Methods]The production characteristics of porcine reproductive and respiratory syndrome virus(CH-1a strain)on Marc-145 clone cells were studied,and the concentration of serum,pH value and harvest technology were optimized,in order to improve the virus titer and increase the yield of virus.[Methods]The production characteristics of porcine reproductive and respiratory syndrome virus(CH-1a strain)on Marc-145 clone cells were studied,and the concentration of serum,pH value and harvest technology were optimized,in order to improve the virus titer and increase the yield of virus.[Results]When the pH of maintenance solution was controlled between 7.8~8.2,the serum-free culture was carried out by whole-change method.The viral titer of the mixed solution was 8.13lgTCID_(50)/ml,and viral load was 1.73×10^(7)copies/mL.[Conclusion]The optimal production technology of porcine reproductive and respiratory syndrome virus(CH-1a strain)on Marc-145 cells was established and the serum-free culture was realized,this study provides a reference for improving the production and quality of inactivated vaccine against porcine reproductive and respiratory syndrome virus(CH-1a strain).