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猪传染性胃肠炎、猪流行性腹泻二联灭活疫苗的工艺摸索

发布时间:2018-05-23 18:47

  本文选题:猪传染性胃肠炎 + 猪流行性腹泻 ; 参考:《江西农业大学》2017年硕士论文


【摘要】:猪传染性胃肠炎(Swine Transmissible Gastroenteritis,TGE)和猪流行性腹泻(Porcine Epidemic Diarrhea,PED)均是由冠状病毒引起,可造成患病仔猪出现呕吐、腹泻及死亡等临床症状,是危害猪群健康的两种重要疫病。由于市场供应的疫苗质量参差不齐,大部分疫苗的中和抗体水平较低,对猪群保护的效果不理想,给养猪业带来了巨大的经济损失。我们通过开展对两种病毒敏感细胞的筛选、病毒的浓缩、免疫佐剂的筛选、免疫增强剂的添加、病毒液保存条件及病毒含量变化实验、肠毒的制备并灭活等实验,对猪传染性胃肠炎、猪流行性腹泻二联灭活疫苗的生产工艺进行了优化并制备了样品,随后,免疫3日龄阴性仔猪后检测其免后两周中和抗体水平。根据本次实验结果,我们总结如下:(1)筛选出较敏感细胞PK-15-1和VERO-1两株,猪传染性胃肠炎和猪流行性腹泻效价TCID50均在10-7/ml以上,且猪传染性胃肠炎合格率90%以上,猪流行性腹泻合格率100%;(2)PEDV制备后一周之内灭活并配苗,TGEV两周之内灭活并配苗能够保证成品质量良好;(3)提高两种病毒的病毒含量,采取对疫苗进行沉淀后除去五分之二上清液的工艺可以提高猪传染性胃肠炎和猪流行性腹泻二联灭活疫苗的中和抗体;(4)相同浓度下,选择颗粒越细,表面积越大的氢氧化铝胶佐剂可以提高猪传染性胃肠炎和猪流行性腹泻两种抗原中和抗体水平。以上实验在一定程度上可达到提高疫苗产品质量和防疫效果的目的。对部分工艺进行固化,从而实现优化猪传染性胃肠炎、猪流行性腹泻二联灭活疫苗工艺的目标。本研究为优化猪传染性胃肠炎、猪流行性腹泻二联灭活疫苗的工艺提供了参考。
[Abstract]:Porcine Epidemic diarrhea and porcine Epidemic diarrhea caused by coronavirus can cause vomiting, diarrhea and death in sick piglets. They are two important infectious diseases which are harmful to the health of pigs. Due to the uneven quality of vaccines supplied in the market, the neutralizing antibody level of most vaccines is low, and the effect of protecting pigs is not ideal, which has brought huge economic losses to the pig industry. We conducted experiments on the screening of sensitive cells for the two viruses, the concentration of viruses, the screening of immune adjuvants, the addition of immune enhancers, the preservation conditions of virus fluids and the changes in virus contents, the preparation and inactivation of enterotoxins, etc. The production process of the inactivated vaccine against infectious gastroenteritis and epidemic diarrhea in pigs was optimized and the samples were prepared. Then the antibody levels were detected after immunizing 3-day-old negative piglets. According to the results of this experiment, we summarized as follows: 1) two sensitive cell strains, PK-15-1 and VERO-1, were screened out. The titers of TCID50 of porcine infectious gastroenteritis and porcine epidemic diarrhea were above 10-7/ml, and the qualified rate of infectious gastroenteritis was over 90%. The qualified rate of porcine epidemic diarrhea was 100%. The virus content of the two viruses was increased by inactivation and inoculation of TGEV within two weeks after the preparation of PEDV. When the supernatant of 2/5 was removed by precipitating the vaccine, the neutralizing antibody of the inactivated vaccine of infectious gastroenteritis and epidemic diarrhea in pigs could be increased at the same concentration, the finer the selected particles were, The larger the surface area of aluminum hydroxide gel adjuvant, the higher the neutralizing and antibody levels of infectious gastroenteritis and epidemic diarrhea. To a certain extent, the above experiments can improve the quality of vaccine products and the effect of epidemic prevention. In order to achieve the goal of optimizing the inactivated vaccine process of infectious gastroenteritis and epidemic diarrhea in pigs, some of the processes were solidified. This study provides a reference for optimizing the inactivated vaccine of infectious gastroenteritis and epidemic diarrhea in pigs.
【学位授予单位】:江西农业大学
【学位级别】:硕士
【学位授予年份】:2017
【分类号】:S858.28

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