肉制品中牛源性成分多重实时荧光PCR检测 方法的建立及初步应用

Establishment and application of multiple real-time fluorescent PCR to detect the beef ingredient in meat products

中文关键词:  多重实时荧光PCR  牛源性成分  肉制品

英文关键词:multiple real-time fluorescent PCR  beef ingredient  meat products

中文摘要:

      目的 建立肉制品中牛源性成分的荧光PCR检测方法。方法 根据牛特异性线粒体DNA片段, 设计合成两对引物, 以生、熟牛肉及超市牛肉加工品为材料, 建立肉制品中牛源性成分的多重实时荧光PCR检测方法, 并用该法与国标法同时对市售的25份肉制品同时进行检测, 通过对其他种类的肉源DNA进行扩增验证方法的特异性; 对含有不同比例牛肉成分的DNA样本进行检测确定检出限。结果 该方法可成功检测出肉制品中的牛源性成分。在25份肉制品检测中, 与国标法检测结果一致。该法的特异性为100%, 灵敏度检测线为1%。结论 本研究成功建立牛源性肉制品的检测方法, 该方法快速简便, 且具有较高的特异性和灵敏度, 可用于市售肉制品中牛源性成分的鉴定。

英文摘要:

      Objective To establish a multiple real-time fluorescent PCR method for the determination of beef ingredient in meat product. Methods Two cattle-specific primers were designed according to the bovine specific mitochondrial DNA fragments, raw, cooked beef and beef processed products were used as materials, multiple real-time fluorescent PCR method was established to detect the beef ingredient in twenty-five beef products, and national standard method was used as positive control. The specificity of the method was verified through the detection of DNA from the meat of other species, and the limit of detection was determined by testing several samples with different proportions of beef. Results The results showed that this method could successfully detect the beef ingredient in meat products. Our results were almost the same as the national standard detection methods in detecting 25 meat samples, with detection rate of 100% and detection limitation of 1%. Conclusion A multiple real-time fluorescent PCR method is successfully established. This method is simple, rapid and has high specificity and sensitivity, which can be applied to identify the beef ingredient in meat products on sales.