为更好地利用实时荧光PCR技术鉴别肉制品的真实属性，创新性地采用微波提取法对样品均质液进行DNA提取，并且根据不同物种动物的基因序列设计特异性的引物与探针，根据实时荧光PCR扩增结果确定肉制品所含物种成分。在30份牛肉制品中，检测出有7份样品含有猪、鸡或鸭源性成分，检出率约为23.3%；在20份羊肉制品中，检测出有5份样品含有猪、鸡或鸭源性成分，检出率为25.0%。微波提取DNA的方法优化了DNA提取流程，使不同类型肉制品中提取DNA可在短时间内完成，极大地提高了检测效率。开发的样品前处理方法、DNA提取方法具有灵敏度高、特异性强等优点，可广泛应用于肉制品及其相关产品真实属性的鉴别，具有良好的社会效益与经济效益。%A unique DNA isolation protocol based on microwave irradiation and density centrifugation for the simultaneous detection of bovine, caprine, ovine, porcine, chicken and duck species in meat products by PCR using newly developed primers was described. The method had been optimized using the primers designed from mitochondrial DNA(mtDNA) sequence and tested in 50 samples. To evaluate the method, 30 beef products and 20 mutton products were analyzed by microwave irradiation and RT-PCR technique. Porcine, chicken and duck content was detected in beef products and mutton products. The design of this protocol makes it possible to pro-cess many samples in a short time. The new set of primers and the method involved showed high sensitivity in detecting beef, mutton, porcine, chicken and duck content in meat products. The present sample preparation method has the potential to be applied to other meat detection systems as well.
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