Instituto de Investigaciones Marinas (IIM-CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain.
Food Chem. 2017 Oct 15;233:182-189. doi: 10.1016/j.foodchem.2017.04.087. Epub 2017 Apr 17.
Species-specific lateral flow dipstick (LFD) assays for the identification of Atlantic cod (Gadus morhua), Pacific cod (Gadus macrocephalus), Alaska pollock (Gadus chalcogrammus) and ling (Molva molva) in food products were developed. The method comprises a PCR system with four sets of specific primers, for each target species. This step was also devised to dual-labeling of PCR products with biotin and 6-FAM, which are then easily read on a lateral flow dipstick, upon which these products are immobilized by a fixed biotin-ligand and visualized with anti-FAM antibody-coated gold nanoparticles. Sensitivity and selectivity were determined for each of the developed assays. Validation of the assays was performed with DNA extracted from commercial fish products, the identification of all samples by PCR-LFD was coherent with the results found with DNA sequencing. Target species were successfully detected in analyzed commercial samples, demonstrating the applicability of this method to the rapid analysis of food products.
开发了用于鉴定食品中大西洋鳕鱼(Gadus morhua)、太平洋鳕鱼(Gadus macrocephalus)、阿拉斯加狭鳕(Gadus chalcogrammus)和鳕鱼(Molva molva)的种特异性侧向流试纸条(LFD)检测方法。该方法包括一个具有四组特定引物的 PCR 系统,用于每个目标物种。该步骤还设计用于对 PCR 产物进行生物素和 6-FAM 的双重标记,然后在侧向流试纸上轻松读取,然后将这些产物通过固定的生物素配体固定,并通过带有抗-FAM 抗体包被的金纳米颗粒可视化。针对每种开发的检测方法确定了敏感性和选择性。通过从商业鱼产品中提取 DNA 对检测方法进行了验证,PCR-LFD 鉴定的所有样品与 DNA 测序结果一致。在分析的商业样品中成功检测到目标物种,证明了该方法在快速分析食品中的适用性。