摘要
为建立兔波氏杆菌(Bb)的快速检测方法,本研究采用纯化重组兔Bb fimN蛋白作为包被抗原建立了间接ELISA抗体检测方法。通过优化反应条件,确定其检测临界OD450nm值为0.295。检测结果显示,该方法对兔Bb标准阳性血清检测的敏感性为1∶216;而与兔巴氏杆菌、兔出血症病毒阳性血清无交叉反应。重复性试验表明,其组内和组间变异系数均小于10%。采用建立的间接ELISA方法对浙江省5家兔场采集的150份兔血清样品的检测阳性率为55%。该方法的建立为进一步研发兔Bb ELISA诊断试剂盒提供了实验依据。
To establish a rapid detection protocol for the antibody detection of Bordetella bronchiseptica in rabbits, the indirect ELISA was developed with the purified recombinant fimN protein as coating antigen. Under the optimized reaction condition, the cut-off value was 0.295. The results showed that the ELISA was specific to detect the antibody against B.bronchiseptica at a sensitivity of 1：216 dilutions for the positive serum, but no cross reaction to the positive sera of Pasteurella multocida and rabbit hemorrhagic disease virus. The coefficients of inter-assay and intra-assay variation were less than 10%. In addition, a total of 150 rabbit serum samples which collected fi＇om five different farms of Zhejiang province were tested by the established method, the seropositive rate was 55%. The rfimN-based ELISA provided a technological basis for the development of the ELISA kit for detection of B. bronchiseptica in fection in rabbits.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第11期899-902,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家现代农业兔产业体系建设疾病预防与控制岗位专家项目(nycytx-44-3-2)
浙江省重大农业项目(2011C12028)
