食品及调味品中罂粟成分的实时荧光PCR检测方法

Detection of Papaver somniferum in Food and Condiment by Real-time PCR

针对食品及调味品非法添加罂粟成分的检测需求,针对罂粟小檗碱桥酶基因设计了TaqMan特异性引物和探针,并建立了食品及调味品样品前处理和DNA提取方法,建立了食品及调味品中罂粟成分TaqMan实时荧光PCR检测方法,方法检出限为0.01%,灵敏度为10pg。与普通PCR法和SYBR Green等荧光染料法相比,TaqMan探针法具有更好的特异性,检出限更低,结果判读直观无污染。方法的建立可作为食品及调味品中非法添加罂粟成分的检测鉴定方法,也可为其他与罂粟相关的方法研究提供借鉴和参考。

A method of real-time PCR is established for detecting Papaver somniferum ingredient in food and condiment. The TaqMan specific primers and probes are designed for Papaver somniferum berberine bridge enzyme bbel. The pre-processing and DNA extraction method is set up for detecting food and condiment samples. The limit of detection （LOD） of Papaver somniferum ingredient in food and condiment for this method is 0.01 %. The absolute sensitivity of Papaver somniferum genomic DNA detection is 10 pg. Compared with the conventional PCR and SYBR Green fluorescent dye PCR such as SYBR Green, TaqMan probe method has better sensitivity and specificity, visible results and without pollution. This method could be applied for detecting Papaver somniferum in food and condiment. Meanwhile, it can provide references for other Papaver somniferum detection methods.