Adenuga Bukola M, Biltes Rita, Villa Caterina, Costa Joana, Spychaj Anita, Montowska Magdalena, Mafra Isabel
LAQV/REQUIMTE, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
Department of Meat Technology, Poznań University of Life Sciences, ul. Wojska Polskiego 31, 60-624 Poznań, Poland.
Foods. 2024 Nov 21;13(23):3728. doi: 10.3390/foods13233728.
Roe deer meat is a prized game product in many European countries. However, concerns exist regarding the accuracy of the amount of declared roe deer in processed game meat foods. This study aimed to develop a reliable method for the detection and quantification of roe deer in commercialized game meat products. A TaqMan probe-based quantitative real-time PCR (qPCR) assay was designed, targeting a single-copy 120-bp region of the roe deer agouti signaling protein (ASIP) encoding gene. The method employed the normalized ∆Cq approach to establish a calibration curve for roe deer detection and quantification within 0.05-50% (/) in complex raw and processed matrices. The method proved to be specific for roe deer identification, achieving limits of detection and quantification of 0.04 ng of roe deer DNA and 0.05% (/) of roe deer in simulated pâté. Following validation with blind samples, highlighting the precision and trueness of the approach, the assay was applied to 46 market samples from four European origins (Poland, Portugal, France, and Spain). The analysis revealed significant discrepancies between declared roe deer content and actual levels in all roe deer labeled products. The global analysis of results, combining the previous survey on red deer species with present roe deer data, identified 61% of mislabeled/adulterated samples due to the absence of deer species, substitution of roe deer with red deer, substitution of fallow deer with other deer species and red deer with pork, and undeclared addition of roe deer. This study demonstrates the effectiveness of the developed qPCR method for accurate roe deer meat authentication in foods, showing its usefulness as a tool for routine food inspection to ensure labeling compliance.
狍肉在许多欧洲国家都是备受珍视的野味产品。然而，人们对加工后的野味肉类食品中所宣称的狍肉含量的准确性存在担忧。本研究旨在开发一种可靠的方法，用于检测和定量商业化野味肉类产品中的狍肉。设计了一种基于TaqMan探针的定量实时PCR（qPCR）检测方法，靶向狍刺鼠信号蛋白（ASIP）编码基因的一个单拷贝120 bp区域。该方法采用标准化的∆Cq方法，在复杂的生基质和加工基质中建立狍肉检测和定量的校准曲线，范围为0.05 - 50%（/）。该方法被证明对狍肉识别具有特异性，在模拟肉酱中实现了0.04 ng狍肉DNA的检测限和0.05%（/）的定量限。在用盲样进行验证后，突出了该方法的精密度和准确性，该检测方法被应用于来自四个欧洲产地（波兰、葡萄牙、法国和西班牙）的46个市场样品。分析显示，所有标记有狍肉的产品中，宣称的狍肉含量与实际水平之间存在显著差异。结合先前对马鹿物种的调查和当前狍肉数据的结果综合分析表明，61%的样品存在错误标注/掺假情况，原因包括未标明鹿的种类、用马鹿替代狍肉、用其他鹿种替代黇鹿以及用猪肉替代马鹿，还有未申报添加狍肉。本研究证明了所开发的qPCR方法在食品中准确鉴定狍肉的有效性，表明其作为确保标签合规的常规食品检测工具的实用性。
