环介导等温扩增技术在西洋参鉴定中的应用

目的 探讨利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)技术鉴别西洋参的方法.方法 针对西洋参的CYP450基因设计特异性引物,应用RT-LAMP(real time-LAMP)、琼脂糖凝胶电泳和可视化检测3种方法对LAMP检测方法进行优化,并对其灵敏性...

Application of Loop-mediated Isothermal Amplification(LAMP)-Based Technology for Authentication of Panax quinuefolium

OBJECTIVE To evaluate a loop-mediated isothermal amplification (LAMP) assay for detection of Panax quinuefolium. METHODS Based on the CYP450 gene of Panax quinuefolium, specific primers were designed. The LAMP assay was optimized by real time-LAMP (RT-LAMP), agarose gel electrophoresis and visual inspection, and its sensitivity and specificity were evaluated. RESULTS The optimal reaction time of LAMP reaction was 60 min, the optimal reaction temperature was 65 ℃, and the optimal ratio of internal and external primers was 8∶1. The sensitivity of the reaction system reached 0.5 ng·μL^-1, and the specificity was good, which could be distinguished from the related plant Panax ginseng. The adulteration behavior in the market was simulated by adding Panax ginseng into Panax quinuefolium, and the detection rate reached 20%. The positive rate of 22 Panax quinuefolium products in the market was 81%.CONCLUSION These results demonstrate that we develop a rapid, simple, specific and sensitive LAMP assay for Panax quinuefolium, which provide a powerful tool for the rapid identification of Panax ginseng and Panax quinuefolium.