中文摘要：食品和饲料产品中转基因生物的定量分析通常需要它们的标签或耐受阈值。基于标准曲线的简单聚合酶链反应（qPCR）是目前流行的技术，但它通常需要与筛选分析相结合。随着世界市场上转基因产品越来越多，qPCR分析变得即费力又昂贵。本文开发并评估了用于多重定量转基因大豆的液滴数字PCR技术。评估的目的是为了便于检验食品和饲料是否符合欧洲联盟现行的转基因生物法规。在欧盟范围内，授权转基因每种原料的标签阈是0.9%；此外，欧盟对某些未经批准的转基因生物设定了0.1%的技术耐受阈。新型多重定量ddPCR技术检测出11种转基因大豆是授权的，其余4种转基因大豆还待欧盟授权。本文还对检测限、定量限、真实性、重复性和鲁棒性等重要参数进行了评估。
外文摘要：Quantification of genetically modified organisms (GMOs) in food and feed products is often required for their labelling or for tolerance thresholds. Standard-curve-based simplex quantitative polymerase chain reaction (qPCR) is the prevailing technology, which is often combined with screening analysis. With the rapidly growing number of GMOs on the world market, qPCR analysis becomes laborious and expensive. Innovative cost-effective approaches are therefore urgently needed. Here, we report the development and inter-laboratory assessment of multiplex assays to quantify GMO soybean using droplet digital PCR (ddPCR). The assays were developed to facilitate testing of foods and feed for compliance with current GMO regulations in the European Union (EU). Within the EU, the threshold for labelling is 0.9% for authorised GMOs per ingredient. Furthermore, the EU has set a technical zero tolerance limit of 0.1% for certain unauthorised GMOs. The novel multiplex ddPCR assays developed target 11 GMO soybean lines that are currently authorised, and four that are tolerated, pending authorisation in the EU. Potential significant improvements in cost efficiency are demonstrated. Performance was assessed for the critical parameters, including limits of detection and quantification, and trueness, repeatability, and robustness. Inter-laboratory performance was also determined on a number of proficiency programme and real-life samples.
作者：Kosir, Alexandra Bogozalec; Spilsberg, Bjorn; Holst-Jensen, Arne; 等.
作者单位：Natl Inst Biol
期刊名称：SCIENTIFIC REPORTS
期刊影响因子：4.259
出版年份：2017
出版刊次：8
点击下载：用于多重定量15种转基因大豆的液滴数字PCR技术的开发与评估研究