Interfibio, Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007 Tarragona, Spain.
Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Naumburger Strasse 96a, 07743 Jena, Germany.
Food Chem. 2019 Jul 30;287:354-362. doi: 10.1016/j.foodchem.2019.02.095. Epub 2019 Feb 28.
In this work, a duplex PCR-Enzyme Linked Oligonucleotide Assay (ELONA) is reported for the sensitive and reliable detection of pork adulteration in beef and chicken products, two of the most widely consumed meat types in the world. The strategy relies on the use of species-specific tailed primers for duplex amplification and simple dilution of the PCR reactions for direct colorimetric detection via hybridization, eliminating the need for any other post-amplification steps. A high sensitivity was achieved, with as low as 71-188 pg of genomic DNA able to be detected using mixtures of control DNA from each species. The strategy was validated using DNA add-mixtures as well as DNA extracted from raw meat mixtures and 0.5-1% w/w pork could be easily detected when mixed with beef or chicken. The proposed approach is simple, sensitive and cost-effective compared to equivalent commercial kits suitable for detecting adulterant pork levels in meat products.
本工作报道了一种用于敏感可靠检测牛肉和鸡肉制品中猪肉掺假的双重聚合酶链反应-酶联寡核苷酸检测（ELONA）方法，牛肉和鸡肉是世界上消费最多的两种肉类。该策略依赖于使用种特异性加尾引物进行双重扩增，并通过杂交直接进行简单稀释的 PCR 反应进行比色检测，无需任何其他扩增后步骤。该方法的灵敏度很高，使用每种物种的对照 DNA 混合物进行检测时，最低可检测到 71-188 pg 基因组 DNA。该策略使用 DNA 添加混合物以及从原始肉混合物中提取的 DNA 进行了验证，当与牛肉或鸡肉混合时，可轻松检测到 0.5-1% w/w 的猪肉。与适用于检测肉类产品中掺假猪肉水平的等效商业试剂盒相比，该方法简单、灵敏且具有成本效益。
