| 豆状带绦虫六钩蚴Tp-dUTPase基因的表达及血清学诊断效果 |
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| 引用本文： | 陈林,;杨德英,;孙家刚,;周英姿,;农向,;谢跃,;古小彬,;杨光友.豆状带绦虫六钩蚴Tp-dUTPase基因的表达及血清学诊断效果[J].兽医大学学报,2014(7):1100-1105. |
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| 作者姓名： | 陈林 ;杨德英 ;孙家刚 ;周英姿 ;农向 ;谢跃 ;古小彬 ;杨光友 |
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| 作者单位： | [1]四川农业大学动物医学院,四川雅安625014; [2]四川省苍溪县畜牧食品局,四川苍溪628400; [3]四川省眉山市畜牧局,四川眉山620032 |
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| 基金项目： | 教育部长江学者和创新团队发展计划资助项目（IRT0848） |
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| 摘 要： | 采用PCR方法从豆状带绦虫六钩蚴中扩增得到Tp-dUTPase基因编码区,构建了pET32a-Tp-dUTPase原核表达载体,以重组蛋白dUTPase作为包被抗原建立的兔豆状囊尾蚴病Dot-ELISA诊断方法对226份兔血清进行了检测。结果显示,Tp-dUTPase基因编码区长447bp,编码148个氨基酸,表达的重组蛋白相对分子质量为36 200,该重组蛋白能与家兔豆状囊尾蚴病阳性血清特异性结合。以重组蛋白Tp-dUTPase作为包被抗原建立的Dot-ELISA诊断方法显示有较高的特异性（85.7%~92.9%）和敏感性（86.4%~88.0%）,与其他种类兔寄生虫病阳性血清有较低的交叉反应。结果表明,纯化的Tp-dUTPase重组蛋白可作为兔豆状囊尾蚴病的诊断抗原。
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| 关 键 词： | 豆状带绦虫 Tp-dUTPase 免疫诊断 Dot-ELISA |
Prokaryotic expression of Tp-dUTPase gene of Taenia pisiformis oncospheres and establishment of dot-ELISA using the expressed protein |
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| Institution: | CHEN Lin, YANG De-ying , SUN Jia-gang, ZHOU Ying-zi , NONG Xiang, XIE Yue, GU Xiao- bin, YANG Guang-you(1. College of Veterinary Medicine, Sichuan Agricultural Universi- ty ,Ya an,Sichuan 625014,Chinas2. Bureau of Animal Husbandry and Food Administration of Cangzi , Cangxi , Sichuan 628400, China 3. Bureau of Animal Husbandry and Food Administra- tion of Meishan ,Meishan, Sichuan 620032, China) |
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| Abstract: | In order to establish diagnostic method of Cysticercus pisiformis,the purified recombinant protein was used for development of an dot-ELISA for detection of anti-Cysticercus pisiformis antibodies.The complete coding region of Tp-dUTPase gene was firstly amplified from the cDNA of activated oncospheres of Taenia pisiformis by RT-PCR.The expression vector of pET32a-Tp-dUTPase was successfully constructed and expressed in Escherichia coli BL21（DE3）.226sera samples of rabbit were tested.The open reading frame of Tp-dUTPase consisted of 447bp and encoding 148amino acids.The recombinant protein was about 36 200.The positive serum of rabbit infceted with Taenia pisiformis larvae could specifically bound to Tp-dUTPase recombinant protein through western blotting.This purified recombinant fusion protein,rTpdUTPase,was probed by dot-ELISA with sera from rabbits infected with Taenia pisiformis larvae and with other parasites.Results showed that the dot-ELISA assay had both high sensitivity（85.7%-92.9%）and specificity（86.4%-88.0%）to detect Taenia pisiformis larvae infections,indicating that it can be used as diagnostic antigen. |
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| Keywords: | Taenia pisiformis Tp-dUTPase gene immune diagnosis dot-ELISA |
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