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| 1型鸭疫里氏杆菌OmpA蛋白间接ELISA方法的建立 | |
| 引用本文： | 孙,郭东春,刘家森,姜骞,司昌德,林欢,崔玉东,曲连东.1型鸭疫里氏杆菌OmpA蛋白间接ELISA方法的建立[J].中国预防兽医学报,2011,33(8). |
| 作者姓名： | 孙 郭东春 刘家森 姜骞 司昌德 林欢 崔玉东 曲连东 |
| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001;黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 2. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨,150001 3. 黑龙江八一农垦大学动物科技学院,黑龙江大庆,163319 |
| 基金项目： | 黑龙江省科技攻关项目(PC09S02) |
| 摘 要： | 为建立检测1型鸭疫里氏杆菌(R.anatipestifer)的间接ELISA方法,本研究根据已发表的R.anatipestifer外膜蛋白A(ompA)基因序列(AF104937)设计引物,扩增1型R.anatipestifer HLG1株的ompA基因,构建重组质粒pHtb-ompA,转化大肠杆菌BL21(DE3),并利用IPTG进行诱导表达。SDS-PAGE和western blot结果表明,表达蛋白约为55 ku,具有良好的抗原活性。以纯化的OmpA为包被抗原建立间接ELISA并对条件进行优化。建立的ELISA具有良好的特异性、敏感性;与HLG1株菌体裂解蛋白为抗原的间接ELISA比较,符合率为91.3%。本研究建立的ELISA方法为R.anatipestifer的流行病学调查和SPF鸭的监测提供了快速、特异的血清学诊断方法。 |
| 关 键 词： | 鸭疫里氏杆菌 OmpA蛋白 间接ELISA |
Development of an indirect ELISA for detection of the antibodies to serotype 1 Riemerella anatipestifer |
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| SUN Yan,GUO Dong-chun,LIU Jia-sen,JIANG Qian,SI Chang-de,LIN Huan,CUI Yu-dong,QU Lian-dong.Development of an indirect ELISA for detection of the antibodies to serotype 1 Riemerella anatipestifer[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(8). | |
| Authors: | SUN Yan GUO Dong-chun LIU Jia-sen JIANG Qian SI Chang-de LIN Huan CUI Yu-dong QU Lian-dong |
| Institution: | SUN Yan1,2,GUO Dong-chun1,LIU Jia-sen1,JIANG Qian1,SI Chang-de1,LIN Huan1,CUI Yu-dong2,QU Lian-dong1*(1.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China,2.College of Animal Science and Veterinary Medicine,Heilongjiang August First Land Reclamation University,Daqing 163319,China) |
| Abstract: | To develop an indirect ELISA assay for detection of antibody against Riemerella anatipestifer,the ompA gene from R.anatipestifer HLG1 strain was amplified using a pair of primers based on the pubished sequence of R.anatipestifer(AF104937) and inserted into pPRO-EX-Htb vector to generate the recombinant plasmid pHtb-ompA for expression of OmpA in E.coli.The result of SDS-PAGE showed that the expressed protein was about 55 ku and western blot results indicated that recombinant protein was recognized with posi... |
| Keywords: | Riemerella anatipestifer OmpA protein indirect ELISA |
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