Sanchiz Africa, Sánchez-Enciso Paulina, Cuadrado Carmen, Linacero Rosario
Food Technology Department, SGIT-INIA, Ctra. La Coruña Km. 7.5, 28040 Madrid, Spain.
Genetics, Physiology and Microbiology Department, Biology Faculty, Complutense University, 28040 Madrid, Spain.
Foods. 2021 Jun 18;10(6):1421. doi: 10.3390/foods10061421.
Peanut () contains allergenic proteins, which make it harmful to the sensitised population. The presence of peanut in foods must be indicated on label, to prevent accidental consumption by allergic population. In this work, we use chloroplast markers for specific detection of peanut by real-time PCR (Polymerase Chain Reaction), in order to increase the assay sensitivity. Binary mixtures of raw and processed peanut flour in wheat were performed at concentrations ranging from 100,000 to 0.1 mg/kg. DNA isolation from peanut, mixtures, and other legumes was carried out following three protocols for obtaining genomic and chloroplast-enrich DNA. Quantity and quality of DNA were evaluated, obtaining better results for protocol 2. Specificity and sensitivity of the method has been assayed with specific primers for three chloroplast markers (mat k, rpl16, and trnH-psbA) and Ara h 6 peanut allergen-coding region was selected as nuclear low-copy target and TaqMan probes. Efficiency and linear correlation of calibration curves were within the adequate ranges. Mat k chloroplast marker yielded the most sensitive and efficient detection for peanut. Moreover, detection of mat K in binary mixtures of processed samples was possible for up to 10 mg/kg even after boiling, and autoclave 121 °C 15 min, with acceptable efficiency and linear correlation. Applicability of the method has been assayed in several commercial food products.
花生（）含有致敏蛋白，这使其对致敏人群有害。食品中花生的存在必须在标签上标明，以防止过敏人群意外食用。在这项工作中，我们使用叶绿体标记物通过实时聚合酶链反应（PCR）对花生进行特异性检测，以提高检测灵敏度。在小麦中对生花生粉和加工花生粉的二元混合物进行了浓度范围为100,000至0.1毫克/千克的检测。按照三种获取基因组和叶绿体富集DNA的方案，从花生、混合物和其他豆类中提取DNA。对DNA的数量和质量进行了评估，方案2获得了更好的结果。使用针对三种叶绿体标记物（mat k、rpl16和trnH-psbA）的特异性引物以及Ara h 花生过敏原编码区域作为核低拷贝靶标和TaqMan探针，对该方法的特异性和灵敏度进行了检测。校准曲线的效率和线性相关性在适当范围内。Mat k叶绿体标记物对花生的检测最为灵敏和有效。此外，即使经过煮沸和121°C 15分钟的高压灭菌，在加工样品的二元混合物中，mat K的检测限仍可达10毫克/千克，效率和线性相关性均可接受。该方法的适用性已在几种商业食品中进行了检测。
