肉制品中马源性成分实时荧光PCR检测方法的建立

Development and application of real-time PCR for identification and detection of horse meat in animal-origin products

目的建立肉制品中马源性成分的实时荧光PCR检测方法。方法参考欧盟实验室推荐的方法设计引物和探针,建立实时荧光PCR检测方法并验证方法的特异性、灵敏度,在北京部分超市、农贸市场、餐馆随机采集国产和进口牛、羊肉制品进行马源性成分检测。结果实时荧光PCR方法对马源性成分具有较高的特异性,与羊、猪、鸡、鸭、兔DNA均无交叉反应,与牛DNA在Ct 33.81出现交叉反应;所建方法对牛、羊肉中人工掺入马肉成分的检测灵敏度是0.1%。对北京市场上随机采集的122份牛、羊肉制品的检测结果显示,所有样品均未检出马肉成分。结论所建肉制品中马源性成分的检测方法简单、特异,检测灵敏度完全达到欧盟对肉类食品中掺入马源性成分比例的要求。

Objective To develop a real-time PCR method for identification and detection of domestic horse meat （Equus caballus） in animal-origin products. Methods The primer and TaqMan-probe was designed and synthesized according to the EU reference laboratory and 87 bp fragments was amplified for horse ingredients. The specificity and sensitivity was tested by artificially spiked horse meat into other domestic meat, such as cattle, sheep, pork, chicken, duck and rabbit. 122 samples of cattle and sheep products were random collected in Beijing market and the detection of horse meat was carried out. Results The real-time PCR in this study has high specificity and sensitivity for horse meat. No cross- reaction was observed between the horse and sheep, pork, chicken, duck and rabbit meat. There was little cross reaction between horse and cattle when the CT value reach 33.81. The method can detect 0. 1% of horse meat mixed with other domestic animal-origin products. No horse meat ingredients were detected in 122 samples in this survey. Conclusion There was no horse meat mixed into cattle and sheep products in Beijing marked.