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当前位置: 首页 > 文章 > 豆状带绦虫Tp18基因的原核表达及其产物的抗原性分析 畜牧兽医学报 2013,44 (11) 1819-1825
Position: Home > Articles > Prokaryotic Expression of Tp18Gene fromTaenia pisiformis and Antigenicity Analysis of the Expressed Product Chinese Journal of Animal and Veterinary Sciences 2013,44 (11) 1819-1825

豆状带绦虫Tp18基因的原核表达及其产物的抗原性分析

作  者：
杨德英;陈林;古小彬;赖松家;孙家刚;杨光友
单  位：
四川农业大学动物医学院;四川省苍溪县畜牧食品局;四川农业大学动物科技学院
关键词：
豆状带绦虫;Tp18基因;原核表达;免疫保护;诊断
摘  要：
为评价豆状带绦虫Tp18重组蛋白对兔豆状囊尾蚴病的免疫保护效果和诊断应用价值,从豆状带绦虫成虫转录组数据库中筛选到Tp18基因,并进行克隆和原核表达,以纯化后的重组蛋白作为免疫原和反应原。免疫保护试验设计了重组蛋白组、佐剂组和PBS组,共免疫2次(间隔14d),二免后的第7天用5 000个豆状带绦虫虫卵攻虫;攻虫后的第49天屠宰实验兔,统计兔体内豆状囊尾蚴的数量。整个试验过程中每周采血,分离血清进行IgG和IgA水平检测。同时,设计了Tp18重组蛋白和PBS重复组,试验程序和检测指标均与第一次试验相同。此外,采用Tp18重组蛋白建立了诊断家兔豆状囊尾蚴病的Dot-ELISA方法,对169份家兔血清进行分析,评估其诊断应用价值。结果表明表达的Tp18基因成熟肽的重组蛋白约为32ku,Western blotting证明该重组蛋白与豆状带绦虫感染兔阳性血清具有良好的反应原性。Tp18重组蛋白组的减虫率为95.59%(重复组为97.38%),极显著高于佐剂组和PBS组(P<0.01);诱导产生的特异性抗体为IgG。以病原学检查为金标准,Tp18重组蛋白建立的DotELISA诊断方法具有较高的敏感性(49/54,90.74%)和特异性(111/115,96.52%)。研究结果表明,Tp18重组蛋白可作为豆状带绦虫的疫苗抗原和诊断抗原,为兔豆状囊尾蚴病的防控提供了理论基础。
译  名：
Prokaryotic Expression of Tp18Gene fromTaenia pisiformis and Antigenicity Analysis of the Expressed Product
作  者：
YANG De-ying;CHEN Lin;GU Xiao-bin;LAI Song-jia;SUN Jia-gang;YANG Guang-you;College of Veterinary Medicine,Sichuan Agricultural University;College of Animal Science & Technology,Sichuan Agricultural University;Animal Husbandry Foodstuff Bureau of Cangxi County;
关键词：
Taenia pisiformis;;Tp18gene;;prokaryotic expression;;immunoprotection;;diagnosis
摘  要：
In order to evaluate the effect of immunoprotection and diagnosis against rabbit Taenia pisiformiscysticercosis by recombinant Tp18(rTp18)protein,Tp18gene was screened from transcriptome of adult T.pisiformis,then was cloned and expressed.r Tp18protein was used as immunogen and atopen.The samples were grouped into three experimental groups,including rTp18protein group,PBS group and adjuvant group.Immunization was performed twice at 14-day intervals.Seven days after the second immunization,each rabbit was experimentally infected orally with 5 000mature viable T.pisiformis eggs,and sacrificed at 49days post-infection.The number of cysticercoids was counted.By the end of the experiment,serum was collected for detection of IgG and IgA every week.Repeated trial of rTp18protein group and PBS group was carried out using identical procedures and conditions.The molecular weight of rTp18protein is about 32kDa.rTp18protein was proved to be well reacted with the positive sera against T.pisiformis from rabbit by Western blotting.Vaccination trials showed that the rTp18protein induced effective immune protection against T.pisiformis cysticercosis with 95.59%reduction(97.38% reduction in repeated trial)in metacestode burdens of vaccinated rabbits,which was significantly higher than PBS and adjuvant group(P<0.01).IgG was the antibody of humoral immunity. Meanwhile,dot-ELISA was used to diagnose T.pisiformis cysticercosis for rabbits.Based on the results of rabbit necropsy of 169abattoir rabbit samples,the relative sensitivity and specificity of the dot-ELISA were 90.74%(49/54)and 96.52%(111/115),respectively.These promising results indicate that rTp18protein is suitable for the development of effective vaccination and diagnositic strategies against cysticercosis in rabbits,providing a foundation for control of T. pisiformis cysticercosis.

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