以BORCT蛋白为包被抗原,优化最佳反应条件,确定阈值并测定其交叉反应,建立了一种特异性检测牛巴贝斯虫抗体的间接ELlSA方法。利用该方法测定了试验感染的2头牛的抗体动态曲线,并对我国四川、甘肃、新疆3个省(自治区)的314份牛血清样品进行了检测。结果显示,经MedCalc12.7.3.0软件分析86份阴性血清和69份阳性血清的检测结果,确定29.69/6抗体比率为该方法的阳性阈值,对应的敏感性和特异性分别为94.2%和96.5%,并且与其他巴贝斯虫、泰勒虫和无浆体阳性血清均无交叉反应。对试验感染动物的检测结果显示,感染后的第6天出现特异性抗体,第12天达到最高值,一直持续到第270天。野外样品检测结果显示,3个省份均有牛巴贝斯虫的分布,平均阳性率为46.82%。
An indirect ELISA for specific detection of Babesia boris antibodies was established using the purified recombinant protein of B. boris rhoptry-associated protein 1 C terminal(BORCT) as a coating antigen by optimizing reaction conditions and evaluating specificity and sensitivity. And then ELISA was used to evaluate antibody kinetics of 2 experimentally infected cattle by B. boris and to test 314 bovine sera collected from Sichuan,Gansu and Xinjiang Provinces. The results showed that the specificity and sensitivi- ty were 96.5% and 94.2% ,respectively,when 29.6% of antibody rate were chosen as a positive threshold via analyzing the data of 86 negative sera and 69 positive sera with MedCalc 12.7.3.0 software. And there were no cross-reactions between BORCT and positive sera of other Babesia, Theileria and Anaplasma spe- cies. Animals experimentally infected with B. boris had sero-converted by 6 days post inoculation and the highest level of antibodies reached after 12 days. The specific antibodies lasted for 270 days post infection. The test for the field samples demonstrated that B. boris was distributed in these provinces and the average positive rate of was 46.82 %.