转基因大豆的PCR-免疫层析筛查方法研究初探

Preliminary Study on PCR-Immunochromatographic Test for Screening Genetically Modified Soybeans

目的建立转基因大豆的PCR-免疫层析(PCR-ICT)快速筛查新技术。方法利用转基因植物通常含有的CaMV35S启动子作为转基因成分的筛查标记,根据其序列设计特异性引物和探针,分别用生物素和地高辛标记,用胶体金免疫层析技术检测并鉴定PCR产物。结果用新建立的PCR-ICT方法可以检出含0.5%转基因大豆的标准品,对大豆样品的检测结果与琼脂糖凝胶电泳检测结果一致。结论PCR-ICT方法通过DNA杂交和金标显色来同时检测、鉴定PCR产物,可以简便、快速地筛查转基因产品。

Objective To develop a PCR-immunochromatographic test （PCR-ICT） for screening genetically modified （GM） soybean. Method The promoter CaMV35S was used as a marker for screening genetically modified organisms （GMOs）. According to the gene sequences of promoter CaMV35S introduced into GMOs, PCR primers were quoted from previous reports and the former primer was labeled with biotin, and a specific probe was designed and labeled with digoxigenin. The PCR amplification product was detected and identified by colloid gold immunochromatographic test. Result 0.5 % of standard GM soybean powder could be detected by the newly developed PCR-ICT. The results of detecting soybean samples using this test consisted with those of using agar gel electrophoresis. Conclusion The newly developed PCR-ICT appears to be a rapid and convenient technique for screening GMOs. It could simultaneously detect and identify specific PCR product by DNA hybridization and colloid gold immunochromatographic test.