基于 TaqMan 探针实时荧光定量 PCR 用于粉丝掺假成分的检测

文章选取红薯、木薯、绿豆、豌豆、马铃薯、玉米6种常见制作粉丝原料的特异性引物和探针，建立实时荧光定量聚合酶链式反应（polymerase chain reaction，PCR）方法用于粉丝中掺假成分的鉴别。水解探针（TaqMan探针）的设计方法是在5'端连接荧光基团羧基荧光素（FAM），3'端连接猝灭基团羧基四甲基罗丹明（TAMAR），当探针发生水解时，荧光基团与淬灭基团分离，荧光基团所产生的荧光即可被仪器检测到。结果表明，该方法的相对灵敏度为1%，绝对灵敏度为 $ 10^{-2} $ ng/ $ \mu $L，可应用于市售60种粉丝中的红薯、木薯、绿豆、豌豆、马铃薯、玉米源性成分的鉴别。基于TaqMan探针实时荧光定量PCR检测方法反应耗时短，具有灵敏度高、特异性好、定量准确等优点，可以作为粉丝中源性成分掺假检测的有效方法之一。

This paper selects specific primers and probes from six common raw materials for making vermicelli, including sweet potato, cassava, mung bean, pea, potato, and corn, to establish a quantitative real-time polymerase chain reaction (RT-qPCR) method for identifying adulterated ingredients in vermicelli. The method of TaqMan probe is to connect the fluorescence group FAM at 5' and the quenching group TAMAR at 3'. When the probe is hydrolyzed, the fluorescence group separates from the quenching group, and the fluorescence generated by the fluorescence group can be detected by the instrument. The results showed that the relative sensitivity of the method was 1%, and the absolute sensitivity was $ 10^{-2} $ ng/ $ \mu $L. It can be applied to the identification of sweet potato, cassava, mung bean, pea, potato and corn-derived components in 60 commercially available vermicelli. The TaqMan probe-based RT-qPCR detection takes a short reaction time, and has the advantages of high sensitivity, excellent specificity, and accurate quantification, making it an effective tool for detecting adulterants in vermicelli.