Q热贝氏柯克斯体间接ELISA检测方法的建立

Establishment of Indirect ELISA Method to Detect Q Fever Coxiella burnetii

本研究以Q热贝氏柯克斯体弱毒株Ⅱ相全菌为包被抗原建立Q热贝氏柯克斯体间接ELISA检测方法,通过方阵滴定法对抗原包被浓度和二抗反应浓度的确定及各项反应条件的优化,确定其阴阳性临界值为0.44。用本方法与IDEXX Q热抗体检测试剂盒对393份牛临床血清进行检测,检出的血清阳性率分别为11.45%和6.10%,符合率为94.66%。结果表明,本试验建立的检测Q热贝氏柯克斯体的间接ELISA法具有较好的特异性及较强的敏感性,可初步应用于Q热贝氏柯克斯体抗体的临床检测。

In this study, the whole bacteria protein of the Coxiella burnetii attenuated strain phase II was used as the antigen and then the indirect ELISA to detect Q fever Co：ciella burnetii was established. Through optimizing the concentration of the coated phase II antigen, the concentration of the second antibody and the reaction condition, we confirmed that the critical value of this method was 0.44. 393 clinical bovine sera were detected by this method and IDEXX Q fever （Coxiella burnetii） Antibody Test Kit. The results showed that the sera positive detection rates were 11.45% and 6.10%, respectively. Compared with the results of the two detection methods, the coincidence was 94.66%. The results demonstrated that sensitivity and specificity of the established indirect ELISA detection method were better and it could be used for the clinical diagnosis of the antibody of Q fever Coxiella burnetii.