[1]孙,郭东春,刘家森,等.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,(08):635-639.
 SUN Yan,GUO Dong-chun,LIU Jia-sen,et al.Development of an indirect ELISA for detection of the antibodies to serotype 1 Riemerella anatipestifer[J].Chinese journal of preventive veterinary medicine,2011,(08):635-639.
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1型鸭疫里氏杆菌OmpA蛋白间接ELISA方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年08期
页码:
635-639
栏目:
诊断和检测技术
出版日期:
2011-08-15

文章信息/Info

Title:
Development of an indirect ELISA for detection of the antibodies to serotype 1 Riemerella anatipestifer
作者:
孙;郭东春;刘家森;姜骞;司昌德;林欢;崔玉东;曲连东;
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;黑龙江八一农垦大学动物科技学院;
Author(s):
SUN Yan12GUO Dong-chun1LIU Jia-sen1JIANG Qian1SI Chang-de1LIN Huan1CUI Yu-dong2QU 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
关键词:
鸭疫里氏杆菌OmpA蛋白间接ELISA
Keywords:
Riemerella anatipestifer OmpA protein indirect ELISA
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。
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 positive serum against R.anatipestifer.An indirect ELISA was developed using the recombinant OmpA as coating antigen.The coincidence of the indirect ELISA coating with recombinat OmpA was 91.3% with the ELISA coating with the lytic bacteria of R.anatipestifer.The indirect ELISA assay with good sensitivity,specificity and repeatability would provide a simple and rapid means for monitoring R.anatipestifer infection and assessing vaccination in the field

参考文献/References:

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备注/Memo

备注/Memo:
基金:黑龙江省科技攻关项目(PC09S02)
更新日期/Last Update: 2011-09-29