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- Title
重组产气荚膜梭菌毒素对家兔的安全性和免疫效力评价.
- Authors
赵晓军; 王美玲; 段跃强; 谭美玲; 石悦萌; 陈千林; 郭庆勇
- Abstract
To investigate the safety and immune efficacy of recombinant Clostridium perfringens a-toxin in rabbits, the recombinant plasmid pVL1393-CPA based on Clostridium perfringens a-toxin gene was constructed in this study, and was co-transfected into Sf9 cells by BD BaculoGold™ kit. The monoclonal recombinant virus was purified by plaque phagocytosis and the target protein of 45ku was expressed in High 5 cells and then detected by SDS-PAGE and western blot. The recombinant a-toxin was inactivated with 0.1 % formaldehyde solution and emulsified with ISA 206 VG adjuvant to prepare the subunit vaccine (antigen content was 20μg/mL). Rabbits (1.5-2.0kg) were injected intramuscularly with the vaccine containing 40μg of the protein, and PBS control group was set up, all experiment rabbits were observed for 7 days post injection. The results showed that the vaccine was safe for rabbits. In another experiment, groups of rabbit (n=10) were immunized twice with 20μg in 21 day's internal. Meanwhile, the adjuvant control group or toxin control group were set up. At 21 days after prime and 14 days after boost, serum was collected for the detection of IgG titers by ELISA. The toxin neutralization test in mice and the protection test in rabbits were used to evaluate the immune protection efficacy of this candidate vaccine. The results showed that the titer of serum IgG antibody in the vaccine immunized group was significantly different from that in the adjuvant control group and the toxin control group (P<0.001). The neutralizing titer of two immunized serum against Clostridium perfringens type A mice was 32, and the rabbits in the vaccine immunized group were healthy after challenge, and the protection rate was 100%. The mortality of rabbits in adjuvant and toxin control groups was 100%. These results indicated that the recombinant Clostridium perfringens a-toxin vaccine prepared in this study has good safety and immune protective efficacy in rabbits, which provides an important reference for the development of other clostridium subunit vaccines.
- Subjects
CLOSTRIDIUM perfringens; NEUTRALIZATION tests; RECOMBINANT viruses; VACCINE effectiveness; CONTROL groups
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2023, Vol 45, Issue 4, p429
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202212021