Abstract:
Foot and Mouth Disease (FMD) is globally pandemic which badly affect the economics of livestock based countries like Pakistan. There are different types of Foot and Mouth Disease Virus (FMDV) among these types O is most prevalent in Pakistan. Recently Pakistan is producing approximately fifteen million doses of non-purified FMD vaccine against the demand of 160 million doses annually. More over the Pakistan is still striving for the development and optimization of concentration as well as purification of FMDV. The present project was designed to develop the technology for the purification of FMDV indigenously. The locally isolated and adapted FMDV type O virus was propagated on adherent culture of BHK-21cells to get final volume of virus one liter. This virus suspension was concentrated by peggylation as well as ultra-filtration method. The purification and quantification of concentrated virus was done by size exclusion chromatography. The results showed that peggylation is better method of concentration up to 603.75 µg/ml with 82.80 % recovery rate than ultra-filtration with 43.90 % followed by chromatography for purification. The PD50 was calculated in bovines at 24, 12, 6, 3 and 1.5 µg of FMDV Ag/dose and it revealed that antigen load of 1.98 µg is the dose, where the 50 % of inoculated animals showed the protective antibody level based upon percent inhibition through antibody detecting ELISA. According to the British pharmacopeia, the vaccine should contain 3PD50 which found equivalent to our findings about 6 µg/dose. The group of animal injected with 6/dose (3.23PD50) showed protective titer up to 20th week post priming.
摘要:
口蹄疫(FMD)是全球流行病,严重影响巴基斯坦等畜牧业国家的经济。在这些O型中,有不同类型的口蹄疫病毒(FMDV)在巴基斯坦最普遍。最近,巴基斯坦正在生产大约1500万剂非纯化口蹄疫疫苗,而每年的需求为1.6亿剂。巴基斯坦的更多地区仍在努力开发和优化FMDV的浓缩和纯化。本项目旨在开发本地纯化FMDV的技术。在BHK-21细胞的贴壁培养物上繁殖局部分离和适应的FMDVO型病毒以获得最终体积的病毒1升。通过聚乙二醇化以及超滤方法浓缩该病毒悬浮液。浓缩病毒的纯化和定量通过尺寸排阻层析进行。结果表明,聚乙二醇化是浓度高达603.75µg/ml的更好方法,回收率为82.80%,优于超滤,43.90%,然后进行色谱纯化。在24、12、6、3和1.5µgFMDVAg/剂量的牛中计算PD50,并显示1.98µg的抗原载量是剂量,其中50%的接种动物通过抗体检测ELISA显示基于抑制百分比的保护性抗体水平。根据英国药典,疫苗应含有3PD50,这相当于我们的发现约6µg/剂。注射6/剂(3.23PD50)的动物组显示保护性滴度直至引发后第20周。