牛副结核分枝杆菌LAMP快速检测方法的建立

Development of a LAMP assay for Mycobacterium avium subsp.Paratuberculosis

为建立牛副结核分枝杆菌(MAP)的快速检测方法,本研究采用环介导等温扩增(LAMP)技术,以MAP的IS900基因序列为靶基因设计特异性引物,建立了MAP特异性的LAMP快速检测方法。经反应条件优化,检测结果显示,建立的LAMP检测方法具有良好的特异性,最低见测量为0.53 pg;与其他病原菌无任何交叉反应。临床样品检测与行业标准的符合率为100%。该LAMP检测方法可以用于MAP的常规检疫。

To establish a quick detection protocol for Mycobacterium avium subsp, paratuberculosis （MAP）, a loop-mediated isothermal amplification （LAMP） assay was developed with primers designed according to the IS900 gene of MAP available in GenBank. Under the optimized amplification conditions. The LAMP assay was specific to MAP with detection limit of 0.53 pg and no cross-reactions with other related bacteria. These results demonstrated that the LAMP assay is a useful technique for routine MAP quarantine.