| 犬瘟热病毒重组核蛋白间接ELISA方法的建立 |
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| 引用本文： | 王琛,袁宝,任文陟,张嘉保.犬瘟热病毒重组核蛋白间接ELISA方法的建立[J].畜牧与兽医,2008,40(5):37-40. |
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| 作者姓名： | 王琛 袁宝 任文陟 张嘉保 |
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| 作者单位： | 1. 吉林大学农学部实验动物中心,吉林,长春,130062;武汉大学生命科学学院,湖北,武汉,430072
2. 吉林大学农学部实验动物中心,吉林,长春,130062 |
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| 基金项目： | 吉林省科技计划发展项目(20071138). |
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| 摘 要： | 将已构建成功的重组质粒pGEX-4T-1-N转化大肠杆菌BL21株,在最佳诱导条件下获得犬瘟热病毒(CDV)重组N蛋白。将表达产物纯化后进行SDS-PAGE和W estern-b lot分析,与CDV标准阳性血清呈阳性反应。本研究初步建立了以纯化的N蛋白为包被抗原的间接ELISA检测方法,经初步试验证实,该方法敏感、特异。试验结果表明,大肠杆菌中表达的CDV N蛋白在免疫原性上与天然核衣壳蛋白具有较高相似性,可作为诊断用抗原。
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| 关 键 词： | 犬瘟热病毒 核衣壳蛋白基因 原核表达 间接ELISA |
| 文章编号： | 0529-5130(2008)05-0037-04 |
| 修稿时间： | 2007年11月28 |
Establishment of an indirect ELISA with a recombinant nucleocapsid protein as antigen for detection of antibodies against canine distemper virus |
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| WANG Chen,YUAN Bao,REN Wen-zhi,ZHANG Jia-bao.Establishment of an indirect ELISA with a recombinant nucleocapsid protein as antigen for detection of antibodies against canine distemper virus[J].Animal Husbandry & Veterinary Medicine,2008,40(5):37-40. |
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| Authors: | WANG Chen YUAN Bao REN Wen-zhi ZHANG Jia-bao |
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| Abstract: | The recombinant plasmid pGEX-4T-1-N was transformed into E.coli BL21 and the recombinant nucleocapsid protein of canine distemper virus(CDV) was obtained under the optimized induction condition.The analysis of SDS-PAGE indicated a protein band with the molecular weight of 55ku.Western-blot analysis showed that the recombinant protein had a positive reaction with standard positive blood serum against CDV.The indirect ELISA method was developed using the purified fusion protein of nucleocapsid as antigen.The preliminary clinical application showed that it is a sensitive and specific method in detecting CDV antibodies,and the coincidence rate between the recombinant protein ELISA and IDEXX CDV kit was up to 96.25%. |
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| Keywords: | canine distemper virus nucleocapsid protein prokaryotic expression indirect ELISA |
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