牛巴贝斯虫GST-MSA-2c融合蛋白抗象间接ELISA检测方法的初步建立

以纯化的牛巴贝斯虫GST-MSA-2c融合蛋白作为检测抗原，通过优化ELISA反应条件，初步建立了检测牛巴贝斯虫血清特异性抗体的间接ELISA方法。方阵试验确定的GST-MSA-2C抗原的最适包被浓度为2.5μg／mL，血清最佳稀释倍数为20倍，ELISA阳性反应的临界值为OD450≥0.347，批内和批间重复试验的变异系数均小于10％。经对多例血清检测表明，所建ELSIA检测法重复性好、特异性强、灵敏度高。

Elementary Development of an Inderect ELISA Assay for Detecting Antibody Against Babesia Bovis GST-MSA-2c Fusion Protein

The purified recombinant fusion protein （GST-MSA-2c） was used as coating antigen,an Indi- rect ELISA assay for the direction of Babesia bovis was elementarily developed by optimizing the reaction conditions. In the cross assay the optimal concentration of coating antigen was 2.5μg/mL and the optimal dilution of serum sample was 1 ： 20. The cutoff value for positive response by established ELISA was OD450nm≥0.347. In the intra-batch and the inter-batch repeated tests, the variation coefficient was less than10%. The results were shown that the ELISA method was highly sensitive, specific and reproducible. This is the first case of using recombinant protein to establish diagnostic method for the ditection of Babesia bovis in China. It was provided a new technical method for the serological dignosis and a large scale of epidemiological investigation on bovine babesiosis.