三重LAMP法检测阿胶中驴、马和猪源性成分

Development of Multiple LAMP Method for Detection of Donkey,Horse and Pig-Derived Ingredients in Asini Corii Colla

为能快速检测阿胶中驴、马、猪源性成分,建立了三重环介导等温扩增(LAMP)检测方法。针对驴、马、猪线粒体基因的保守区域,设计并筛选出3套LAMP特异性引物。通过优化反应条件,建立三重LAMP方法,将检测为阳性的产物进行酶切法鉴定,判断样本中驴、马或猪源性成分。驴、马和猪的检测限分别为7.9×10^(-3)、1.36×10^(-3)ng和8.5×10^(-2)ng。结果表明该检测方法操作简便、特异性强、灵敏度高,适合于市场阿胶真伪鉴别与驴、马和猪源性成分的鉴定。

A method of multiple loop-mediated isothermal amplification(LAMP)for detection donkey,horse and pig-derived ingredients in Asini corii Colla was developed.Three sets of specific primers were designed and screened for the conserved regions of mitochondrial genes of donkey,horse and pig.After optimizing the reaction conditions,the method of multiple LAMP was established.And the positive products were identified as the donkey,horse or pig-derived ingredients in the samples by restriction analysis.The limits of detection for donkey,horse and pig DNA in Asini corii Colla were 7.9×10^(-3),1.36×10^(-3)ng and 8.5×10^(-2)ng,respectively.The results showed multiple LAMP has the advantages of simple operation,specificity and excellent sensitivity,which can be applied in the identification of donkey,horse and pig-derived ingredients in Asini corii Colla.