Spice adulteration not only seriously interferes with their flavoring functions but also leads to life-threatening poisoning for consumers. To overcome the limitations of traditional methods in spice adulteration detection, a multiplex allele-specific PCR system was developed for molecular discrimination of four commonly used spices, Foeniculum vulgare Mill., Zanthoxylum bungeanum Maxim., Illicium verum Hook.f., and Syzygium aromaticum (L.) Merr. & L.M.Perry, from their corresponding adulterants based on chloroplast SNP markers. The developed assay, eliminating the obstacles of DNA sequencing and false negative results, can detect 0.1% of spice adulteration down to 0.01 ng level of genomic DNA with absolute allelic specificity and favorable efficiency. Based on the results, a standard operating procedure for using multiplex allele-specific PCR for spice adulteration detection was established. Therefore, the present study provided a simple, reliable, and sensitive molecular method for adulteration detection of spices.

香料掺假不仅严重干扰其调味功能，还会导致消费者中毒而危及生命。为了克服传统方法在香料掺假检测中的局限性，开发了多重等位基因特异性PCR系统，对四种常用香料（茴香、花椒、八角茴香和丁香）进行分子鉴别。(L.)梅尔。& LMPerry，根据叶绿体 SNP 标记从相应的掺假品中提取。所开发的检测方法消除了DNA测序和假阴性结果的障碍，可以检测0.1%的香料掺假，低至0.01 ng基因组DNA水平，具有绝对的等位基因特异性和良好的效率。根据结果​​，建立了使用多重等位基因特异性 PCR 检测香料掺假的标准操作程序。因此，本研究为香料掺假检测提供了一种简单、可靠、灵敏的分子方法。