C型产气荚膜梭菌类毒素疫苗与抗毒素血清的制备研究

发布时间：2018-05-17 09:26

本文选题：C型产气荚膜梭菌类毒素疫苗 + 怀孕母猪　；参考：《山东农业大学》2015年硕士论文

【摘要】：为制备具有高效保护力的C型产气荚膜梭菌类毒素疫苗,利用C型产气荚膜梭菌标准菌株(NCTC 3180)所产外毒素经甲醛灭活并加入佐剂乳化后,制备了C型产气荚膜梭菌类毒素疫苗;将该疫苗免疫母猪后检测母猪初乳和仔猪血清的抗体效价;用制备的C型产气荚膜梭菌类毒素疫苗分别多次免疫猪,隔段时间后采集血液,制备血清,测定血清中的抗毒素效价。本研究分为3部分进行:第一部分:C型产气荚膜梭菌类毒素疫苗的制备。对C型产气荚膜梭菌标准菌种(NCTC3180)进行复苏、增菌、产毒,再用0.22μm滤器过滤除去菌体得到毒素溶液,然后用甲醛对毒素进行灭活后,分别加入白油佐剂和氢氧化铝胶佐剂制得两种C型产气荚膜梭菌类毒素疫苗。第二部分:类毒素疫苗的检验。参照《中华人民共和国兽用生物制品质量标准》分别对疫苗进行了安全性检验,无菌检验,免疫效力检验。分别在分娩前30和15天免疫接种怀孕母猪,通过间接ELISA和毒素中和试验评估母猪初乳和14日龄仔猪血液中抗体效价水平。结果显示,母猪初乳和仔猪血清中抗毒素效价分别为1:3200,1:1600,母猪初乳和仔猪血液中的中和效价均为1:6。表明制备的类毒素疫苗效价高并安全可靠,免疫实验动物后能对仔猪起到一定的保护作用。第三部分:抗毒素血清的制备。采用制备的C型产气荚膜梭菌白油佐剂类毒素疫苗多次免疫猪,采集血液,制备血清,参照《中华人民共和国兽用生物制品质量标准》分别对血清进行了安全性检验和无菌检验;通过间接ELISA法和小鼠的血清中和试验测定血清效价。结果显示:血清的安全性检验和无菌检验均符合标准;间接ELISA法测定猪血清中的抗α抗体效价为1:6400;间接ELISA法测定猪血清中的抗β抗体效价为1:12800。0.1m L的C型产气荚膜梭菌高免血清可以中和500个MLD。
[Abstract]:In order to prepare Clostridium perfringens type C toxoid vaccine with high protective power, the toxin produced by Clostridium perfringens strain NCTC 3180 was inactivated by formaldehyde and emulsified with adjuvant, then the toxin vaccine of Clostridium perfringens type C was prepared. After immunizing sows, the antibody titers of sows colostrum and piglets serum were detected, the pigs were immunized with the C type Clostridium perfringens toxoid vaccine for several times, and the blood samples were collected at intervals to prepare serum. The antitoxin titers in serum were determined. This study is divided into three parts: the first part is the preparation of Clostridium perfringens toxoid vaccine. The standard strain of Clostridium perfringens type C, NCTC3180), was resuscitated, increased bacteria, produced toxin, filtered the bacteria with 0.22 渭 m filter to get toxin solution, then inactivated the toxin with formaldehyde. Two C-type Clostridium perfringens toxoid vaccines were prepared by adding white oil adjuvant and aluminum hydroxide adjuvant respectively. The second part: the examination of toxoid vaccine. With reference to the quality Standard of Veterinary Biological products of the people's Republic of China, the safety, aseptic and immunological efficacy of the vaccine were tested. Pregnant sows were immunized 30 and 15 days before delivery. The antibody titers in colostrum and 14 day piglets were evaluated by indirect ELISA and toxin neutralization tests. The results showed that the antitoxin titers in serum of sows and piglets were 1: 3 200 and 1: 1600, respectively, and the neutralization titers in colostrum and blood of sows were 1: 6, respectively. The results showed that the prepared toxoid vaccine had high titer, safety and reliability, and could protect piglets after immunizing experimental animals. Part three: preparation of antitoxin serum. Pigs were immunized with C type Clostridium perfringens white oil adjuvant toxoid vaccine for many times. Blood was collected and serum was prepared. According to the quality standard of animal biological products of the people's Republic of China, the safety test and aseptic test of serum were carried out, and the titer of serum was determined by indirect ELISA method and serum neutralization test of mice. The results showed that the safety test and aseptic test of serum were in accordance with the standard. The titer of anti 伪 antibody in pig serum was 1: 6400 by indirect ELISA method, and 500 MLDs could be neutralized by indirect ELISA assay when the titer of anti 尾 antibody in swine serum was 1: 12800. 1m L and that of Clostridium perfringens type C high immune serum was 1: 6400.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.4

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