China Medical University, Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, Taichung, 40402, Taiwan.
National Pingtung University of Science and Technology, Department of Food Science, Pingtung, 91201, Taiwan.
Sci Rep. 2017 Mar 14;7:44238. doi: 10.1038/srep44238.
Plant-based food ingredients such as garlic, Chinese leek, Chinese onion, green onion and onion are widely used in many cuisines around the world. However, these ingredients known as the "five forbidden vegetables" (FFVs) are not allowed in some vegetarian diets. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for the detection of FFVs using five respective LAMP primer sets. The specific primers targeted the ITS1-5.8S-ITS2 nuclear ribosomal DNA sequence regions among the five vegetables. The results demonstrated that the identification of FFVs using the newly developed LAMP assay is more sensitive than the traditional PCR method. Using pepper, basil, parsley, chili and ginger as references, established LAMP primer sets showed high specificity for the identification of the FFV species. Moreover, when FFVs were mixed with other plant ingredients at different ratios (100:0, 50:50, 20:80, 10:90, 5:95, 2:98, and 1:99), no cross-reactivity was evident using LAMP. Finally, genomic DNAs extracted from boiled and steamed FFVs in processed foods were used as templates; the performance of the LAMP reaction was not influenced using validated LAMP primers. Not only can FFV ingredients be identified but commercial foods containing FFVs can also be authenticated. This LAMP method will be useful for the authentication of FFVs in practical food markets in the future.
植物性食品成分，如大蒜、韭菜、葱、洋葱和大葱，广泛应用于世界各地的许多菜肴中。然而，这些被称为“五辛菜”（FFV）的食材在某些素食饮食中是不被允许的。在这项研究中，开发了一种环介导等温扩增（LAMP）检测方法，使用五个特定的 LAMP 引物对 FFV 进行检测。特异性引物针对这五种蔬菜的 ITS1-5.8S-ITS2 核核糖体 DNA 序列区域。结果表明，与传统 PCR 方法相比，新开发的 LAMP 检测方法对 FFV 的鉴定更敏感。使用辣椒、罗勒、欧芹、辣椒和姜作为参考，建立的 LAMP 引物对显示出对 FFV 物种鉴定的高度特异性。此外，当 FFV 与其他植物成分以不同比例（100:0、50:50、20:80、10:90、5:95、2:98 和 1:99）混合时，LAMP 没有明显的交叉反应。最后，使用验证过的 LAMP 引物，从加工食品中煮沸和蒸煮的 FFV 提取的基因组 DNA 作为模板，LAMP 反应的性能不受影响。不仅可以鉴定 FFV 成分，还可以鉴定含有 FFV 的商业食品。这种 LAMP 方法将有助于未来在实际食品市场中对 FFV 进行鉴定。
