四重PCR结合全自动电泳系统检测食品转基因成分

Detection of genetically modified food by quadruplex PCR and automated electrophoresis system

根据转基因食品中最常见的四种外源元件(CaMV35S启动子、NOS终止子、cry1Ab/cry1Ac基因、bar基因)的核苷酸序列,设计特异性检测引物,扩增产物大小分别为101、180、301、430bp,通过引物浓度、退火温度的优化,特异性、灵敏度测试,并结合微流体芯片全自动电泳技术,建立了同时检测这4个参数的四重PCR方法。结果表明,该方法可特异性地从复杂样品中检测出预期转基因成分,检测灵敏度达到0.1%。本方法特异性好,灵敏度高,适用于食品中转基因成分的快速筛查。

A novel quadruplex PCR screening approach combined with the automatic electrophoresis technology for genetically modify food was developed.This assay was consisted of four PCR systems targeting on four genetic elements widely introduced into GMOs, such as CaMV35 S promoter, NOS terminator, cry] Ab/cryl Ac gene and bar gene.The amplified product by quadruplex PCR assay was 101,180,301,430bp, respectively.The multiplex PCR conditions were optimized according to the primer concentration and annealing temperature.Then, the specificity and sensitivity of the established quadruple PCR method was tested. Results showed that this quadruple PCR approach can specifically detect the targeted exogenous genetically modified ingredients from complex food samples,with the relative limit of detection of OA%. In conclusion, the multiplex PCR method offering highly specificity and sensitivity was suit to the rapid screening detection for genetically modified components in food samples.