牛巴贝斯虫GST-MSA-2c融合蛋白间接ELISA血清学诊断方法的建立

摘要

[目的与方法]在优化表达牛巴贝斯虫GST-MSA-2c融合蛋白和建立ELISA反应条件的基础上,进-步探讨牛巴贝斯虫GST-MSA-2c融合蛋白及其所建立的ELISA检测方法的特异性,从而建立牛巴贝斯虫GST-MSA-2c融合蛋白间接ELISA血清学检测方法.[结果]牛巴贝斯虫GST-MSA-2c融合蛋白间接ELISA检测方法能排除GST的干扰,与其它梨形虫病无交叉反应,与牛巴贝斯虫巢式PCR检测方法的阳性符合率为96%.[结论]所建立的GST-MSA-2c融合蛋白间接ELISA血清学检测方法重复性好、特异性强、灵敏度高.这是国内首次利用重组蛋白抗原建立的牛巴贝斯病血清学诊断方法,为大规模地进行牛巴贝斯虫病的流行病学调查和血清学诊断提供有效的技术手段.%[ Objective and Method ] The optimal exprssion of CST - MSA - 2C fusion protein and reaction conditions of CST - MSA - 2c indirect ELISA assay were studied in the previous paper. The purposes of this paper were to further discuss the specificity of CST - MSA - 2C fusion protein and GST - MSA - 2c indirect ELISA assay and to establish the standard procedure for this method . [ Result] The positive interference of murine GST antiserum and nested PCR could be eliminated and no cross reactions were found among; the piroplasms. The coincidence precentage of identified positive serum samples for bovine piroplasmosis in indirect MSA - 2c ELISA were 96％.. [ Conclusion]The established CST - MSA - 2c indirect ELISA obtained highly sensitive, specific and reproducible characteristics .which characteristics was the first case to establish an indirect MSA - 2c ELISA assay method for detection of specific antibody against Babesia bovis in China. This test provided a new effective technical method for the large - scale epidemiological survey and serological dignosis of bovine babesiosis infected with Babesia bovis .