实时荧光PCR法检测明胶的牛、猪源性成分

Origin determination of bovine and porcine gelatin by Real-time PCR

为了快速准确鉴别明胶的动物源性,根据线粒体DNA基因COX I(cytochrome coxidase subunit I)的种间多态性差异合成特异性引物及探针,通过实时荧光PCR技术扩增与识别特异性基因片段。运用明胶牛、猪源性成分的荧光PCR检测方法并进行特异性、检出限、重复性实验。结果显示:该方法能够快速准确检测明胶的动物源性,特异性强、灵敏度高、重复性好,对掺杂的牛源性或猪源性成分的检出限均为0.1%,克服了明胶黏度高、不易溶解,多重加工工艺对检测结果干扰严重的难点。实时荧光PCR法为检测明胶的动物源性成分提供了有效手段,保障食品添加剂的质量与安全。

In order to quickly and accurately identify the origin for gelatin,primers and probes were synthesized based on characteristic sequence of COX I(cytochrome coxidase subunit I)of mitochondrial DNA in pork and beef.Real-time fluorescent PCR was used to amplify and identify specific gene sequences to establish a method for determination of the origin of gelation..Fluorescent PCR method,detection limit and reproducibility were studied.Result shows that the method is highly specific,sensitive and reproducible,it is simple to operate.The detection limits for both bovine and porcine products were 0.1%.This method overcomes gelatin’s high viscosity and insolubility as well as interference factors due to multi-steps processing in gel making.Real-time fluorescent PCR provides an effective way of detecting origins of gelatin and ensures the quality and safety of gelatin product.