摘要
采用十二烷基肌氨酸钠法提取哈氏弧菌(Vibrio harveyi)SpGY020601株的外膜蛋白(OMPC),采用酚水法提取溶藻弧菌(Vibrio alginolyticus)EpGS021001株的脂多糖(LPS)。通过碳化二亚铵(EDC)介导的缩合反应,将哈氏弧菌的OMPC与溶藻弧菌的LPS偶联。OMPC-LPS偶联物、未偶联的哈氏弧菌OMPC、溶藻弧菌LPS、哈氏弧菌OMPC和溶藻弧菌LPS的简单混合物以及生理盐水按相同程序免疫卵形鲳(Trachinotus cvatus)。检测血清溶菌酶活性、血清抗体微量凝集反应结果显示,卵型鲳经OMPC-LPS偶联物、OMPC、LPS以及二者的简单混合物免疫后,各免疫组间血清溶菌酶活性没有显著差异(P>0.05),但均显著高于生理盐水对照组(P<0.05);OMPC-LPS偶联物免疫组的血清抗溶藻弧菌EpGS021001抗体效价较单纯溶藻弧菌LPS免疫组、简单混合物免疫组出现得早,抗体滴度高;与此类似,OMPC-LPS偶联组中抗哈氏弧菌SpGY020601的血清抗体效价较单纯哈氏弧菌OMPC组、简单混合组出现得早,抗体滴度高,且持续时间长;对EpGS021001、SpGY020601的攻击,OMPC-LPS偶联物免疫组的保护率分别为85%和95%,高于单纯溶藻弧菌LPS免疫组的70%、单纯哈氏弧菌OMPC免疫组的75%,也高于简单混合物免疫组的80%和82.5%。
Vibrio alginolyticus and V.harveyi,the potentially pathogenic bacteria in southern China,caused tremendous economical loss for marine fish culture.To prevent the disease caused by V.alginolyticus and V.harveyi at the same time,a conjugate vaccine was developed.Lipopolysaccharide(LPS) was extracted from V.alginolyticus,and outer membrane protein complex(OMPC) was extracted from V.harveyi.Then LPS was conjugated to OMPC by 1-ethyl-3(3-dimethylaminopropyl) carbodiimide(EDC) mediated condensation.During the experimental periods,Trachinotus cvatus were immunized with the OMPC-LPS conjugates,unconjugated LPS,unconjugated OMPC,simple mixture of LPS and OMPC,and physiological salt solution(control) in the same procedure respectively.The results showed that there were no significant differences in lysozyme activity detected among the four vaccine groups(P>0.05),but remarkable differences were observed between the vaccine groups and control group(P<0.05).Compared with the anti-EpGS021001 antibodie,it appeared earlier and its titer higher in OMPC-LPS conjugates group than in unconjugated LPS and the simple mixture groups(P<0.05).Also,the anti-SpGY020601 antibodies in OMPC-LPS conjugates group appeared earlier with higher antibodies titers and lasted longer than those in unconjugated OMPC and the simple mixture groups(P<0.05).In addition,protective effects of OMPC-LPS conjugates,unconjugated LPS,unconjugated OMPC,and simple mixture groups were evaluated.After challenged with EpGS021001 and SpGY020601,the RPS of OMPC-LPS conjugates group was 85% and 95% respectively,which was significantly higher than the groups of unconjugated LPS(70%) and unconjugated OMPC(75%),so did higher than that of the simple mixture groups(80% and 82.5%).
出处
《中国水产科学》
CAS
CSCD
北大核心
2007年第6期990-995,共6页
Journal of Fishery Sciences of China
基金
国家863计划"海水养殖鱼类重要病原菌高效疫苗研制"(2006AA100308)
广东省科技计划"海水养殖鱼类重大疾病疫苗及快速诊断"(2004A20401001)
关键词
溶藻弧菌
哈氏弧菌
外膜蛋白
脂多糖
偶联疫苗
Vibrio alginolyticus
Vibrio harveyi
lipopolysaccharide
outer membrane protein complex
conjugated vaccine
