肉制品中植源性转基因成分多重荧光定量PCR检测方法的建立

摘要

为改善产品品质,肉制品加工过程中常常添加植物源性成分,当前转基因农作物的商品化及其在市场上的广泛流通导致肉制品中被带入植源性转基因成分的风险增加.以转基因植物中常涉及的调控元件CaMV 35S启动子、NOS终止子以及标记基因NPTⅡ为检测目标,设计相应的引物和Ta q man探针,利用载体pRⅠ101-AN DNA为模板,通过优化反应体系和反应参数,建立肉制品中植源性转基因成分的单重和多重荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测方法.通过比较分析,考察多重荧光定量PCR检测方法的灵敏性、重复性和准确性,结果表明,多重荧光定量PCR检测方法灵敏度高、重复性好且与单重体系的检测结果具有很好的一致性.%Plant-derived components are often added to meat products during processing to improve their quality. The commercialization and wide circulation in the market of genetically modified crops bring increasing risk of introducing genetically modified components into meat products. In this study, a multiplex fluorescence quantitative polymerase chain reaction (PCR) assay was described for detecting added genetically modified ingredient derived from plants in meat products.Specific primers and Ta q man probes targeting the CaMV 35S promoter, NOS terminator and neomycin phosphotransferase Ⅱ (NPTⅡ) marker gene were designed and the pRⅠ 101-AN DNA vector was used as template. The sensitivity, repeatability and accuracy of the multiplex PCR assay were confirmed by comparison with traditional PCR. Consistent results were obtained using the two methods.