College of Biological and Pharmaceutical Engineering, Jilin Agricultural Science and Technology University, Jilin, Jilin 132101, China.
Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun, Jilin 130033, China.
Food Chem. 2020 Jul 15;318:126541. doi: 10.1016/j.foodchem.2020.126541. Epub 2020 Mar 2.
In recent years, adulteration of meat and meat products has become a major food safety issue. PCR and real-time PCR technologies are mainstream methods used to identify animal-derived components. However, these technologies rely highly on costly equipment and professional technicians; they are therefore difficult to use in resource-limited settings. In this study, a novel, highly sensitive molecular assay, Pig-PCR-Strip (Pig specific polymerase chain reaction-Lateral flow strip), was developed for rapid detection of pig and swine-derived components. The assay is based on PCR amplification, hybridization of the PCR product to the probe, followed by detection using a strip format. Using this format, the PCR product can be detected by the naked eye within 3 min, and provides a basis for the migration of species-specific detection to a point-of-care (POC) microfluidic format. The Pig-PCR-Strip can detect pork components at a concentration of 0.01% in adulterated meat, and the limit of detection is up to 10 fg of target DNA. The assay was specific to pork and did not cross-react with other non-target species. It also can be used for commercial samples and complex food samples. It is a promising new tool for detection of pig-derived meat and can be rapidly modified for identifying other species. It could be widely used for solving problems related to meat quality assurance, species authentication, and traceability.
近年来，肉类及其制品掺假已成为主要的食品安全问题。聚合酶链式反应（PCR）和实时 PCR 技术是用于鉴定动物源性成分的主流方法。然而，这些技术高度依赖昂贵的设备和专业技术人员，因此在资源有限的环境中难以使用。本研究开发了一种新颖的、高灵敏度的分子检测方法——猪 PCR 条带法（Pig-specific polymerase chain reaction-Lateral flow strip，Pig-PCR-Strip），用于快速检测猪肉和猪源性成分。该检测方法基于 PCR 扩增、PCR 产物与探针的杂交，然后使用条带格式进行检测。使用这种格式，PCR 产物可在 3 分钟内用肉眼检测到，并为将物种特异性检测迁移到即时检测（point-of-care，POC）微流控格式提供了基础。猪 PCR 条带法可在掺杂的肉类中检测到浓度为 0.01%的猪肉成分，检测限可达 10 fg 目标 DNA。该检测方法对猪肉具有特异性，与其他非目标物种无交叉反应。它还可用于商业样本和复杂食品样本。这是一种用于检测猪源性肉类的有前途的新工具，可快速修改用于鉴定其他物种。它可广泛用于解决与肉类质量保证、物种鉴定和可追溯性相关的问题。
