基于贝氏柯克斯体急性期抗原A的间接ELISA方法建立及应用

Establishment of an indirect ELISA method based on Coxialla burnetii recombinant AdaA and its application

为开发快速有效的Q热血清学诊断技术,以贝氏柯克斯体急性期抗原A(acute disease antigen A,AdaA)为诊断靶标分子,构建了基于该重组抗原的间接ELISA方法,完成了来自福建某牛场的200份临床牛血清样本检测,并对检测结果进行了评估。结果表明:以商品化ELISA试剂盒ID Screen■Q热为金标准,建立的间接ELISA检测特异性为94.4%(152/161),检测敏感性为84.6%(33/39),与感染牛的常见病原体如衣原体、布氏杆菌、副结核杆菌无交叉。应用该诊断方法检测发现,福建某牛场约有21%(42/200)的牛为Q热阳性。该结果证实,基于AdaA的间接ELISA方法具有较高的特异性及敏感性,有良好的应用价值,可以用于牛Q热的血清学监测。

The aim of this study was to develop a rapid and effective diagnostic system for prevention and control of Q fever.A candidate antigen called acute disease antigen A(acute disease antigen A,AdaA)of Coxialla burnetii underwent recombinant expression and was used as a diagnostic target to construct an indirect ELISA.A total of 200 clinical serum samples from a cattle farm in Fujian Province,China,were detected by this assay,and the results of the assay were evaluated.The results showed that,compared with the commercial kit ID Screen■Q fever used as a standard,the indirect ELISA established in this study possessed detective specificity and sensitivity of 94.4%(152/161)and 84.6%(33/39),respectively.Cross-reactivity of the system was also evaluated,and it was found to be highly specific to Q fever.All the chlamydiosis-,brucellosis-and paratuberculosis-confirmed cattle samples were negative.By using this system,up to 21%(42/200)of the animal serum samples in this flock were positive for Q fever antibodies.The present indirect ELISA was proved to possess good detection specificity and sensitivity,indicating a promising application value in cattle Q fever surveillance programs.