狂犬病口服疫苗的诱饵制作及免疫中和抗体检测

发布时间：2019-01-07 11:52

【摘要】：狂犬病又名恐水症,即民间所说的疯狗病,是由狂犬病病毒引起的以感染中枢神经为特征的一种人畜共患的传染病。狂犬病病毒能感染所有的温血动物,一经感染发病,几乎100%死亡,是目前已知的死亡率最高的传染病之一。据世界卫生组织调查,目前全世界每年死于狂犬病的人数多达55000人,而绝大多数发生在发展中国家,并且主要发生在不发达的农村和山区。目前防治狂犬病的最有效的方法是在暴露后进行高免血清封闭以及免疫狂犬病疫苗,人用的狂犬病疫苗基本都是灭活苗,而动物使用的疫苗主要是弱毒苗,也有少量是进口的灭活苗。由于注射疫苗价格较高,注射不方便等缺点给防治和消灭狂犬病带来了极大的困难。本课题在以前实验室研究的口服疫苗基础之上,根据国外发达国家野外投放口服疫苗的经验,依据犬的生物学特性,利用广西现有的经济资源研制了多种狂犬病口服疫苗诱饵。通过口服途径免疫犬,在第0天、14天、28天、42天、56天和70天采集血液分离血清,用酶联免疫吸附试验(ELISA)和快速免疫荧光抑制实验(RFFIT)检测了犬免疫口服疫苗后中和抗体水平,验证了本实验室研制的口服疫苗的效果。ELISA检测结果显示通过口服疫苗免疫的犬产生了中和抗体,抗体水平0.5IU/ml,产生了免疫保护作用；通过RFFIT试验,犬只也检测到了中和抗体,中和抗体水平0.5IU/ml,两次免疫组中和抗体水平非常接近0.5IU/ml,跟阳性对照组结果基本一致,说明口服疫苗对实验动物具有一定的保护作用。ELISA和RFFIT检测这两种方法实验结果存在一定的差异,需要进一步的研究。本次实验结果也显示了口服疫苗是有一定的免疫效果的,需要进一步改进提高疫苗质量。总之,本实验证明了狂犬病口服疫苗的可行性,为进一步推广和应用口服疫苗免疫野生动物、流浪犬、家犬打下了一定的基础。
[Abstract]:Rabies, also known as hydrophobia, is a zoonotic disease caused by rabies virus. Rabies virus can infect all warm-blooded animals. According to the World Health Organization, the number of people who die of rabies every year in the world is as high as 55000, and the vast majority of the deaths occur in developing countries, and mainly in the underdeveloped rural and mountainous areas. At present, the most effective way to prevent and cure rabies is to block the high immune serum after exposure and to immunize rabies vaccine. The rabies vaccine used in human is basically inactivated vaccine, but the vaccine used in animals is mainly attenuated vaccine. There are also a small number of imported inactivated seedlings. Because of the high price and inconvenient injection, it is difficult to prevent and eliminate rabies. On the basis of the oral vaccine previously studied in laboratory, according to the experience of foreign developed countries in field administration of oral vaccine, according to the biological characteristics of dogs, a variety of rabies oral vaccine decoys were developed by using the existing economic resources in Guangxi. Dogs were immunized orally, and blood serum was collected on day 0, day 14, day 28, day 42, day 56 and day 70. (ELISA) and (RFFIT) were used to detect the neutralizing antibody level of canine oral vaccine. The results of ELISA test showed that the dogs immunized with oral vaccine produced neutralizing antibody, and the antibody level was 0.5 IU / ml, which resulted in immune protection. Through the RFFIT test, neutralizing antibodies were also detected in dogs. Neutralizing antibody levels were 0.5 IU / ml. The neutralization antibody level in the two immunization groups was very close to 0.5 IUP / ml, which was basically the same as that in the positive control group. The results show that oral vaccine has certain protective effect on experimental animals. There are some differences between ELISA and RFFIT test results, which need further study. The results also show that oral vaccine is immune to a certain extent, need to further improve the quality of the vaccine. In conclusion, this experiment proved the feasibility of oral rabies vaccine, which laid a foundation for the further promotion and application of oral vaccine against wild animals, stray dogs and domestic dogs.
【学位授予单位】：广西大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.1

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