Alonso-Rebollo Alba, Ramos-Gómez Sonia, Busto María D, Ortega Natividad
Department of Biotechnology and Food Science, University of Burgos, Plaza Misael Bañuelos, s/n, 09001 Burgos, Spain.
Department of Biotechnology and Food Science, University of Burgos, Plaza Misael Bañuelos, s/n, 09001 Burgos, Spain.
Food Chem. 2017 Oct 1;232:827-835. doi: 10.1016/j.foodchem.2017.04.078. Epub 2017 Apr 13.
The applicability of qPCR in olive-oil authentication depends on the DNA obtained from the oils and the amplification primers. Therefore, four olive-specific amplification systems based on the trnL gene were designed (A-, B-, C- and D-trnL systems). The qPCR conditions, primer concentration and annealing temperature, were optimized. The systems were tested for efficiency and sensitivity to select the most suitable for olive oil authentication. The selected system (D-trnL) demonstrated specificity toward olive in contrast to other oleaginous species (canola, soybean, sunflower, maize, peanut and coconut) and showed high sensitivity in a broad linear dynamic range (LOD and LOQ: 500ng - 0.0625pg). This qPCR system enabled detection, with high sensitivity and specificity, of olive DNA isolated from oils processed in different ways, establishing it as an efficient method for the authentication of olive oil regardless of its category.
定量聚合酶链反应（qPCR）在橄榄油认证中的适用性取决于从油中获得的DNA和扩增引物。因此，设计了四种基于trnL基因的橄榄特异性扩增系统（A-、B-、C-和D-trnL系统）。对qPCR条件、引物浓度和退火温度进行了优化。对这些系统的效率和灵敏度进行了测试，以选择最适合橄榄油认证的系统。所选系统（D-trnL）与其他含油物种（油菜、大豆、向日葵、玉米、花生和椰子）相比，对橄榄具有特异性，并且在较宽的线性动态范围内表现出高灵敏度（检测限和定量限：500ng - 0.0625pg）。该qPCR系统能够以高灵敏度和特异性检测从不同加工方式的油中分离出的橄榄DNA，使其成为一种高效的橄榄油认证方法，无论其类别如何。
