摘要
从健康中华鳖血清中提取抗体并用Sephadex及DEAE 5 2进行了纯化 ,再免疫新西兰白兔制备兔抗鳖抗体的抗血清 ,纯化后以辣根过氧化物酶 (HRP)标记酶标复合物 ,方阵滴定法确定酶标复合物的最适稀释度为 1/ 32 0 0。用温和气单胞菌TL970 42 4株制成铝胶佐剂灭活苗免疫健康中华鳖 ,经一次免疫和二次免疫后 ,分别在不同时间采集血清 ,用ELISA法测定血清抗体变化规律 ,结果表明中华鳖对该菌苗产生良好的体液免疫应答 ,免疫后 2 0d抗体水平达到高峰 ,在测定期间内二次免疫组的抗体水平高于一次免疫组 ,表明中华鳖存在着二次免疫应答 ,但与哺乳类相比 ,中华鳖的二次应答则相对较弱。免疫后 90d对一次免疫组的鳖进行了攻毒试验 ,10d后免疫鳖 10 0 %存活 ,而非免疫对照组鳖则 10 0 %死亡 ,表明温和气单胞菌TL970 42 4株灭活苗免疫效果良好。
An enzyme linked-immunosorbent assay(ELISA)for detection of Aeromonas sobria (As)antibody in Chinese soft-shelled turtle (Trionyx sinensis,Ts)was developed. Chinese soft-shelled turtle antibody was purified by sephadex and DEAE-52.Then inoculated into the New Zealand white rabbit with Freund complete adjuvant.Rabbit anti-turtle IgG was also purified and coagulated with Horseradish peroxidase(HRP).It was determined by square titration method that a dilution of 1∶3200 of this enzyme complex was optimal in terms of test sensitivity.Immune serum was collected at different time after the turtles were immunized with Aeromonas sobria(As)-TL970424 inactived vaccine once or twice.ELISA methods were applied to detecting the change of antibody in serum.The result indicated that Chinese soft-shelled turtle could mount good humoral immune response as well as the second immune response to As inactived vaccine.However this second immune response is weak when compared to that of mammals.Challenge test was carried out 90 days after immunization.No mortality was recorded with turtles injected once whereas there was 100%mortality from group of non-immunized turtles.This result indicated that As inactived vaccine was effective in protecting immunized turtles from attack of this bacteria within 3 monthes post immunization.
出处
《水生生物学报》
CAS
CSCD
北大核心
2003年第1期27-30,共4页
Acta Hydrobiologica Sinica
基金
浙江省自然科学基金 ( 399439)资助项目
