【摘要】 目的研究冻干保护剂中明胶浓度对布鲁杆菌活疫苗活菌率的影响,为布病疫苗保护剂中最佳明胶浓度的确定提供理论依据。方法将布鲁杆菌活疫苗接种到代肝琼脂培养基,转种,扩大培养,用含明胶浓度分别为0%、0.25%、0.5%、1%的4种保护剂收获。经浓度测定,调配浓度,冷冻干燥,封口等工序,制备1 000亿/m L和5亿/m L两种浓度的布鲁杆菌活疫苗,测定冻干后布病疫苗的活菌数。4℃保存样品冻干封口后2 d、2周、1个月、3个月测定活菌数。37℃保存样品冻干封口后2周,1个月测定活菌数。结果冻干后2 d含不同明胶浓度的1000亿/ml疫苗活菌率水平较接近,但冻干后3个月,含1.0%明胶保护剂疫苗活菌率降低比例明显高于其他明胶含量的疫苗。5亿/ml的低浓度疫苗,随着明胶浓度增高,其活菌率降低幅度明显增加,冻干后2 d不含明胶保护剂疫苗活菌率为65.24%,而含1.0%明胶保护剂疫苗活菌率仅为13.34%。含1.0%明胶保护剂疫苗稳定性也较差,冻干后4℃保存3个月的疫苗活菌率降低比例为60.94%,冻干后37℃保存2周的疫苗活菌率降低比例为78.41%。结论冻干保护剂中明胶的浓度愈高,布病疫苗的活菌率愈低,且稳定性越差。更多还原

【Abstract】 Objective To study the effect of gelatin concentration of cryoprotectant on living bacteria rate of Brucella vaccine,and provide theoretical basis for the determination of the best gelatin concentration.Methods The Brucella vaccine was inoculated into the liver agar medium,subcultivation and enlarge cultivation were conducted,results were collected with protective agents of contain gelatin concentration of 0%,0.25%,0.5% and 1% respectively.After concentration determination,allocate concentration,freeze drying,the sealing process and other procedures,100 billion / m L and 500 million / m L of concentration of Brucella vaccine were prepared,the number of living bacteria after freeze drying was determined.Living bacterium numbers were determined after freeze drying and sealing for 2 days,2 weeks,1 month and 3 months at 4 ℃.As well as the numbers after freeze drying and sealing for 2 weeks and 1 month.Results The living bacteria rate levels of 100 billion / m L vaccine with different gelatin concentration 2 days after freeze drying were similar,but 3 months after freeze drying,the reduce rate of the living bacteria rate of vaccine with 1.0% gelatin protective agent is significantly higher than that of other gelatin concentration.500 million / ml low concentration of vaccine,along with the higher concentration of gelatin,the living bacterium rate reduction increased significantly,the living bacteria rate of vaccine with gelatin- free 2 days after freeze drying was 65.24%,but that of vaccine with1.0% gelatin protective agent was 13.34%.The stability of vaccine with 1.0% gelatin protective agent was also poor,the reduce rate of the living bacteria rate of 4 ℃ saving vaccine 3 months after freeze drying was 60.94%,the reduce rate of the living bacteria rate of 37 ℃ saving vaccine 2 weeks after freeze drying was 78.41%.Conclusion The higher the concentration of gelatin in cryoprotectant,the lower the rate of live bacteria vaccine,and the worse the stability.更多还原