五重聚合酶链反应作为食品中水母种类鉴定的筛选检测方法。

Pentaplex PCR as screening assay for jellyfish species identification in food products.

机构信息

FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.

出版信息

J Agric Food Chem. 2014 Dec 17;62(50):12134-43. doi: 10.1021/jf504654b. Epub 2014 Dec 3.

Abstract

Salted jellyfish, a traditional food in Asian Countries, is nowadays spreading on the Western markets. In this work, we developed a Pentaplex PCR for the identification of five edible species (Nemopilema nomurai, Rhopilema esculentum, Rhizostoma pulmo, Pelagia noctiluca, and Cotylorhiza tuberculata), which cannot be identified by a mere visual inspection in jellyfish products sold as food. A common degenerated forward primer and five specie-specific reverse primers were designed to amplify COI gene regions of different lengths. Another primer pair targeted the 28SrRNA gene and was intended as common positive reaction control. Considering the high level of degradation in the DNA extracted from acidified and salted products, the maximum length of the amplicons was set at 200 bp. The PCR was developed using 66 reference DNA samples. It gave successful amplifications in 85.4% of 48 ready to eat products (REs) and in 60% of 30 classical salted products (CPs) collected on the market.

摘要

海蜇，一种亚洲国家的传统食品，如今正在西方市场上流行起来。在这项工作中，我们开发了一种五重 PCR 方法，用于鉴定五种可食用物种（沙海蜇、黄斑海蜇、海月水母、夜光藻和口冠海蜇），这些物种在作为食品销售的海蜇产品中仅凭肉眼无法识别。设计了一个通用的退化正向引物和五个种特异性的反向引物，以扩增 COI 基因区域的不同长度。另一对引物针对 28SrRNA 基因，作为通用阳性反应对照。考虑到从酸化和盐渍产品中提取的 DNA 降解程度很高，将扩增子的最大长度设定为 200bp。该 PCR 使用 66 个参考 DNA 样本进行开发。它成功地扩增了 85.4%的 48 种即食产品（REs）和 60%的 30 种市场上收集的传统盐渍产品（CPs）。