National Reference Laboratory for GM Food and Feed, GMO Unit, Istituto Zooprofilattico Sperimentale del Lazio e della Toscana "Mariano Aleandri", Rome, Italy.
Igiene delle tecnologie alimentari e dell'alimentazione animale, Istituto Zooprofilattico Sperimentale dell'Abruzzo e Molise "G. Caporale", Teramo, Italy.
J Sci Food Agric. 2020 Mar 30;100(5):2121-2129. doi: 10.1002/jsfa.10235. Epub 2020 Jan 17.
The number and variety of genetically modified organisms (GMOs) used globally for the production of food and feed, and potentially circulating in the European Union (EU), is constantly increasing. This implies an additional effort for the EU enforcement laboratories to optimize available resources, to contain costs and time. A well established approach for streamlining the analytical workflow is the introduction of a screening step, typically based on a smart set of real-time polymerase chain reaction (PCR) screening methods. The multiplexing strategy, allowing the detection of several screening elements simultaneously, is a further optimization of this step.
In this study, we present the validation of a real-time PCR duplex assay for the pat and bar screening elements to be easily incorporated in the GMO diagnostic routine. We also provide a comparison between this method and the related singleplex and pre-spotted assays.
Our results fully respect all the validation parameters suggested by the Minimum Performance Criteria of the European Network of GMO Laboratories. Furthermore, the duplex assay is equivalent in terms of performance compared to the other two methods, but it shows a higher overall flexibility and cost effectiveness. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
用于生产食品和饲料的、并且可能在欧盟（EU）流通的遗传修饰生物体（GMO）的数量和种类正在不断增加。这意味着欧盟执法实验室需要额外努力来优化现有资源，以控制成本和时间。一种简化分析工作流程的成熟方法是引入筛选步骤，通常基于一组智能实时聚合酶链反应（PCR）筛选方法。多重策略允许同时检测几个筛选要素，是该步骤的进一步优化。
在这项研究中，我们验证了一种用于 pat 和 bar 筛选要素的实时 PCR 双重检测方法，以便于纳入 GMO 诊断常规。我们还比较了这种方法与相关的单重和预点样检测方法。
我们的结果完全符合欧洲 GMO 实验室网络的最低性能标准中建议的所有验证参数。此外，与其他两种方法相比，双重检测在性能上是等效的，但它显示出更高的整体灵活性和成本效益。 © 2019 作者。John Wiley & Sons Ltd 代表化学工业协会出版的《食品科学与农业杂志》。
