牛布鲁菌荧光标记免疫层析试纸条快速检测方法的建立

Establishment of Fluorescence Labeled Immunochromatographic Strip Method for Rapid Detection of Bovine Brucellosis

为建立一种快速诊断牛布鲁菌病的方法,以原核表达、纯化的外膜蛋白OMP22作为抗原,以带荧光标记的IgG抗体作为标记物制备免疫层析试纸条,建立一种布鲁菌病快速检测方法,并对该方法的敏感性和特异性进行验证。结果显示,制备的试纸条最低可检出1∶100稀释的牛布鲁菌标准阳性血清,且与牛结核、牛蓝舌病、牛病毒性腹泻、牛口蹄疫、牛巴氏杆菌病、牛白血病的血清无交叉反应;试纸条检测结果与商品化ELISA检测试剂盒(IDEXX)的符合率为93.5%。以上结果表明,建立的荧光标记免疫层析试纸条方法能快速检出牛血清中的布鲁菌抗体,具有良好的敏感性和特异性,是一种适合现场初筛的检测方法。

To establish a new method of rapidly detecting Brucella ,the rocombiant outer membrane protein OMP22 was expressed in Escherichia coli and purified to serve as detection antigens. Immunochromato- graphic strips were prepared with fluorescent-labeled IgG antibody as the marker, and the sensitivity and specificity of the method were analyzed.The minimum number of fluorescent-labeled immunechromatogra- phy strip established can be detected at 1 ＂ 100 times diluted bovine brucellosis standard positive serum, and there is no cross reaction with the sera of other related diseases.The rate of concordance between strips and commercial ELISA kit （IDEXX） was 93.5%.It is indicated that he fluorescent-labeled immunochromatographic test strips can be used to detect brucellosis antibodies in bovine serum rapidly with good sensitivity and specificity,and it has great potential in preliminary detection of the antibody of bovine brucellosis serum quickly in practice.