论文部分内容阅读
目的建立以重组外膜蛋白为基础的钩端螺旋体抗体间接ELISA检测方法。方法以基因重组技术获取重组钩端螺旋体外膜蛋白LipL32,以该蛋白为抗原,特异的钩体抗血清进行ELISA方阵滴定、交叉性试验、阻断试验,并对北京地区的70份犬血清使用建立的ELISA方法以及德国Virion公司的全菌体钩端螺旋体ELISA试剂盒进行相互验证。结果方阵滴定试验确立以100ng/孔为抗原包被浓度,1∶160为血清稀释度。交叉性试验具有广泛性、阻断试验标明该方法特异性强、灵敏度高。两种方法数据经χ2检验,两者检出率之间差异不显著。结论重组LipL32蛋白具有结合活性。初步建立了以重组LipL32蛋白为抗原的钩端螺旋体抗体间接ELISA检测方法。
Objective To establish an indirect ELISA method for the detection of Leptospira antibody based on recombinant outer membrane protein. Methods Recombinant Leptospira interrogans lipoprotein LipL32 was obtained by gene recombination technique. The protein was used as antigens and specific antiserum to do ELISA titration, crossover test and blocking test, and 70 serum samples from Beijing The established ELISA method and the whole German company Virion’s Leptospira ELISA kit for mutual validation. Results square matrix titration test to 100 ng / hole as antigen coating concentration, 1:160 serum dilution. Cross-testing is extensive, blocking test shows that the method is specific and sensitive. Two methods of data by χ2 test, the difference between the detection rate was not significant. Conclusion Recombinant LipL32 protein has binding activity. An indirect ELISA method for detecting Leptospira antibody using recombinant LipL32 protein as an antigen was initially established.