A novel fusion protein-based indirect enzyme-linked immunosorbent assay for the detection of bovine tuberculosis

文献类型： 外文期刊

第一作者： Liu, Siguo

作者： Liu, Siguo;Guo, Sheping;Wang, Chunlai;Shao, Meili;Zhang, Xiuhua;Guo, Yang;Gong, Qiang

作者机构：

关键词： fusion protein;bovine tuberculosis;indirect ELISA

期刊名称：TUBERCULOSIS （ 影响因子：3.131； 五年影响因子：2.966 ）

ISSN： 1472-9792

年卷期： 2007 年 87 卷 3 期

页码：

收录情况： SCI

摘要： Enzyme-linked immunosorbent assay (ELISA) for diagnosis of bovine tuberculosis has been widely explored over the years. Three Mycobacterium bovis-specific antigen genes, namely, mpb70, mpb83, and esat-6 were recombined in tandem by spliced overlap extension technology and expressed in Escherichia coli to obtain the fusion protein (rM7083-E6). Western blot analysis showed that rM70-83-E6 can specifically react with bovine tuberculosis-positive sera but not those from cattle infected with other bovine diseases such as bovine paratuberculosis. An indirect ELISA (iELISA) method was established with rM70-83-E6 as the diagnostic antigen. The diagnostic criteria were determined using 150 serum samples from healthy cattle. Analyses of 85 serum samples from cattle with bovine tuberculosis and 100 serum samples from healthy cattle demonstrated that the sensitivity of the iELISAwas 69.4% (59/85) and the specificity was 96.0% (96/100). Moreover, 46 out of 67 purified protein derivative (PPD) skin test-positive samples were also positive by iELISA, giving a positive coincidence of 68.7%, while all 50 PPD skin test-negative samples were negative by iELISA, giving a negative coincidence of 100%. The total coincidence between iELISA and the PPD skin test was 82.1%. This study demonstrated that iELISA using rM70-83E6 antigen is simple, sensitive and easy to perform and can be used to analysis of a large number of samples for serodiagnosis of bovine tubercuiosis. (C) 2006 Elsevier Ltd. All rights reserved.

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