米饭和米酒中Bt基因快速检测方法的建立

Rapid Detection of Bt Gene in Cooked Rice and Rice Wine

建立米饭和米酒中Bt基因环介导等温扩增技术(LAMP)检测方法。以抗虫Bt基因(GeneBank登录号Y09787)为研究对象,利用LAMP技术,针对Bt基因的6个区域设计4条特异性引物,并对镁离子浓度、Bst DNA聚合酶添加量、反应温度、反应时间等扩增条件及体系的特异性、检测灵敏度进行探索。结果表明:成功建立了米饭和米酒的LAMP定性检测方法。所建立的方法能够简便、快速、有效的检测米饭和米酒中的Bt基因,检测限为0.005%。该LAMP检测方法具有高度的特异性及灵敏度,不需要特殊仪器,结果可靠。

We developed a loop-mediated isothermal amplification （LAMP） method for the detection ofBt gene in cooked rice and rice wine. Bt gene （GeneBank accession number Y09787） was amplified by a set of four special primers that were designed according to six regions of the target gene. We optimized the reaction conditions Mg2＋ concentration, Bst DNA polymerase concentration, temperature and time. We also explored the specificity and sensitivity of the reaction. The results showed that the LAMP reaction system allowed easy and effective detection of Bt gene with a high specificity and sensitivity. The limit of detection was 0.005%. The LAMP detection method is specific, sensitive and reliable without need for special instruments.