实时浊度LAMP法检测豆制品中转基因成分

LAMP (Loop-mediated Isothermal Amplification) Detection of Genetically Modified Soybean Products

通过设计转基因大豆内源基因(Lectin)、外源基因NOS和2个转基因大豆品系(GTS 40-3-2,MON89788)特异性LAMP引物,建立快速检测转基因大豆以及鉴定转基因大豆品系的LAMP体系,并选取国内豆制品利用CTAB法进行DNA提取,并针对上述基因进行PCR和LAMP扩增的对比,建立起了检测大豆制品中转基因成分的定性的LAMP方法。结果表明:4个LAMP方法均有较好的特异性,除了NOS、GTS 40-3-2的检测限为0.1%外,其他LAMP方法的灵敏度为0.01%,均能满足实际检测要求;在实时浊度LAMP法检测大豆制品转基因成分的鉴定中,传统的CTAB法能成功提取大豆粗加工制品的DNA,且DNA纯度能够进行核酸扩增反应。利用转基因外引物进行的普通PCR结果与实时浊度LAMP结果是一致的,说明建立的转基因实时浊度LAMP检测方法有很好的准确性。

Specific LAMP primers were designed on the basis of published sequence of lectin gene, NOS termi- nator and two GM soybean strain GTS40-3-2 and MON89788. CTAB DNA extraction method was used to extract DNA of soybean products from local supermarkets. Conventional PCR and LAMP assays were conducted to verify the accuracy of the LAMP method. The results showed that the LAMP assays with the primers designed in the study are specific and sensitive. Traditional CTAB method can successfully extract DNA from soybean products and the DNA purity is suitable for nucleic acid amplification. The PCR results were consistent with LAMP results. This indicated that LAMP method is very accurate.