Ready-to-use single tube quadruplex PCR for differential identification of mutton, chicken, pork and beef in processed meat samples

A reliable and sensitive identification method is required to tackle food adulteration mainly in meat production. We developed a dry reagent based ready-to-use single tube quadruplex PCR assay for accurate identification of chicken, mutton, beef and pork. The assay was found to be specific and reproducible. Thermo-stability studies of lyophilized PCR master mix were conducted at different temperature and time intervals, which revealed significant stability for 75 days at 4°C and for 60 days at 25°C. The developed assay was shown to be sensitive down to 16 pg DNA per reaction and the detection limit was found to be 0.01% (w/w) of each species. Furthermore, this method has been applied to the analysis of 68 commercial meat products and the results indicated that nine samples contained non-declared meat components. This dry reagent-based quadruplex PCR assay can be utilized to monitor various processed food products and also to maintain quality control in food industries mainly in the resource-limited settings.

为应对主要发生在肉类加工环节的食品掺假问题，亟需开发可靠且灵敏的鉴定方法。本研究开发了一种基于干试剂的即用型单管四重聚合酶链式反应（Polymerase Chain Reaction，PCR）检测方法，可精准鉴定鸡肉、羊肉、牛肉及猪肉成分。该检测方法具备良好的特异性与可重复性。针对冻干PCR预混液开展了不同温度与时间梯度的热稳定性研究，结果显示其在4℃条件下可稳定保存75天，25℃条件下可稳定保存60天。该方法的检测灵敏度可达每反应体系16皮克DNA，检出限为各目标物种成分占比0.01%（质量分数）。此外，本方法已应用于68款市售肉类制品的检测分析，结果显示其中9份样品含有未标识的肉类成分。该基于干试剂的四重PCR检测方法可用于各类加工食品的质量监测，尤其适用于资源受限场景下的食品行业质量管控工作。