实时荧光定量PCR检测羊肉制品中鼠源性成分

目的建立基于实时荧光PCR技术的肉制品中鼠源性成分的快速检测方法。方法以羊和鼠的细胞色素b基因序列设计特异性引物和Taqman荧光探针,通过特异性、灵敏性及模拟混合肉样检测实验,建立羊肉制品中鼠源性成分实时荧光定量PCR检测方法。结果该检测方法具有良好的特异性和灵敏度,在50mg羊肉和鼠肉的混合样品检测中,鼠源性成分检测限可低至1%。结论所建立的鼠源性成分检测的实时荧光定量PCR方法,为肉制品质量控制提供了有效的技术手段,弥补了利用RTi-PCR检测肉制品中鼠源性成分的技术空白。

Detection of mouse-derived ingredients in mutton products by Taqman real-time PCR

Objective To establish a rapid method for detecting mouse-derived ingredients from mutton products using the real-time fluorescence quantitative PCR technology. Methods Specific primers and Taqman probes were designed to target the conserved regions of cytochrome b（ cyt b） gene of sheep and mouse. The specificity of the established method was evaluated by direct DNA amplification with various meat samples,while the sensitivity were tested with meat mixtures containing 10%,5%,2% and 1% of mouse meat,respectively. Results The assay was highly specific and sensitive,and the detection limit of mouse-derived ingredients was as low as 1% in 50 mg mixture of mutton and mouse meat. Conclusion The Taqman real-time PCR protocol established by this study provides a reliable means for quality control of meat products,and fills in the technology gap in RTi-PCR detection of mouse-derived ingredients.