摘要
针对药食同源物种藏红花及其常见掺伪品红花、胡萝卜、莲和玉米构建基于TaqMan探针的实时荧光PCR法。根据藏红花和红花的核糖体内转录间隔区(internal transcribed spacer,ITS)基因设计特异性引物和探针,并结合已有研究中胡萝卜、莲和玉米的特异性引物探针对藏红花及其掺伪品进行实时荧光PCR检测。对新设计的藏红花和红花引物探针特异性和检出限进行验证。结果显示,本研究建立的基于TaqMan探针的实时荧光PCR法具有良好的特异性,能将藏红花与其他掺伪品区分。检出限可达到0.1%(w/w)。对市售的20个样品进行实时荧光PCR法和已有标准中的分子生物学检测法检测,发现存在藏红花及红花样品掺伪现象。以上研究结果说明,本研究方法可应用于藏红花的真伪检测,为监管部门提供技术支持。
A real-time fluorescence PCR method based on TaqMan probes was established to identify the authenticity of medicinal and edible saffron and its common adulterants,including safflower,carrot,lotus,and maize.Specific primers and probes were designed based on the conserved internal transcribed spacer(ITS)gene of saffron and safflower.Combined with previously reported specific primers and probes for carrot,lotus,and corn,real-time fluorescent PCR assays were conducted to detect saffron and its adulterants.The specificity and detection limit of the newly designed primers and probes for saffron and safflower were validated.The results showed that the TaqMan probe-based real-time fluorescent PCR method established in this study exhibited high specificity,effectively distinguishing saffron from its adulterants.The detection limit achieved was as low as 0.1%(w/w).Twenty commercial samples were tested by this real-time PCR method and molecular biological detection methods from existing standards,revealing adulteration in both saffron and safflower samples.With its strong specificity and high sensitivity,this real-time fluorescence PCR method enables rapid and accurate authentication of saffron products,providing crucial technical support for market supervision.
作者
朱成杰
王坤
余灏
张倩
赵星宇
王远洋
任佳影
赖心田
ZHU Chengjie;WANG Kun;YU Hao;ZHANG Qian;ZHAO Xingyu;WANG Yuanyang;REN Jiaying;LAI Xintian(Shenzhen Academy of Metrology and Quality Inspection,Shenzhen 518131;East China University of Science and Technology,Shanghai 200237)
出处
《中国食品添加剂》
2025年第7期195-205,共11页
China Food Additives
