高敏荧光免疫层析分析法检测牛分枝杆菌抗体
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S855.3

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新疆生产建设兵团重点领域科技攻关计划项目(2020AB015)


Study on the Application of the Hypersensitive Fluorescence Immunoassay in Detecting Antibodies against Mycobacterium bovis
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    摘要:

    为探究高敏荧光免疫层析分析法检测牛分枝杆菌(M. bovis)抗体的可行性,对该方法的特异性、灵敏性、稳定性及临床适用性进行了分析。特异性试验结果显示,该方法能够检出M. bovis抗体阳性血清参考品,而对布鲁氏菌阳性血清、O型口蹄疫阳性血清、A型口蹄疫阳性血清、样品稀释液、结核菌素皮内变态反应(TST)阴性牛血清样品等均为阴性;灵敏性试验结果显示,该方法最低可检测到3 200倍稀释的牛结核(bTB)强阳性血清参考品;稳定性试验结果显示,该方法检测bTB强阳性血清和阳性血清的变异系数分别为5.48%和5.83%;临床样本检测结果显示,该方法与IDEXX M. bovis ELISA抗体检测试剂盒的符合率为86.36%,具有较高的一致性,且差异不显著。以上结果表明,高敏荧光免疫层析分析法特异性强、灵敏度高、结果可靠,可作为TST检疫的补充抗体检测方法,用于bTB的诊断、检疫和净化。

    Abstract:

    In order to verify the feasibility of the hypersensitive fluorescence immunoassay for detection of antibodies against Mycobacterium bovis(M. bovis),the specificity,sensitivity,stability and clinical applicability of the method were analyzed. It was found that,by specificity test,the method could be used to detect positive serum reference materials of antibodies against M. bovis,but failed for the positive serums of Brucella,foot-and-mouth disease type O(FMD-O)and type A(FMD-A),sample diluent,and bovine negative serums of tuberculin skin test(TST),etc;by sensitivity test,strong positive serum reference materials of bovine tuberculosis(bTB)3 200 times diluted could be still detected;by stability test,the coefficients of variation(CV)of strong positive serum and positive serum were 5.48% and 5.83%,respectively;and by clinical sample test,the coincidence rate of the established method and IDEXX M. bovis ELISA kit was 86.36%,with high consistency and not significant difference. In conclusion,the hypersensitive fluorescence immunoassay was with strong specificity,sensitivity and reliable results,which could be used for diagnosis,inspection,quarantine and purification of bTB as a supplementary test of TST.

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张家瑞,屈勇刚,樊晓旭,张力,剡文亮,孙淑芳,李敏,岳建国,李岩.高敏荧光免疫层析分析法检测牛分枝杆菌抗体[J].《中国动物检疫》编辑部,2022,39(11):121-125.

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