RIKILT Wageningen UR, P.O. Box 230, 6700 AE, Wageningen, The Netherlands.
Anal Bioanal Chem. 2014 Feb;406(5):1397-410. doi: 10.1007/s00216-013-7562-1. Epub 2013 Dec 20.
The ever-increasing production of genetically modified crops generates a demand for high-throughput DNA-based methods for the enforcement of genetically modified organisms (GMO) labelling requirements. The application of standard real-time PCR will become increasingly costly with the growth of the number of GMOs that is potentially present in an individual sample. The present work presents the results of an innovative approach in genetically modified crops analysis by DNA based methods, which is the use of a microfluidic dynamic array as a high throughput multi-detection system. In order to evaluate the system, six test samples with an increasing degree of complexity were prepared, preamplified and subsequently analysed in the Fluidigm system. Twenty-eight assays targeting different DNA elements, GM events and species-specific reference genes were used in the experiment. The large majority of the assays tested presented expected results. The power of low level detection was assessed and elements present at concentrations as low as 0.06 % were successfully detected. The approach proposed in this work presents the Fluidigm system as a suitable and promising platform for GMO multi-detection.
不断增加的转基因作物生产对高通量基于 DNA 的方法提出了要求，以执行转基因生物 (GMO) 的标签要求。随着单个样本中可能存在的 GMO 数量的增长，标准实时 PCR 的应用将变得越来越昂贵。本工作介绍了一种基于 DNA 的方法进行转基因作物分析的创新方法，即将微流控动态阵列用作高通量多检测系统。为了评估该系统，制备了六个具有递增复杂性的测试样品，进行预扩增，然后在 Fluidigm 系统中进行分析。实验中使用了 28 种针对不同 DNA 元件、GM 事件和种特异性参考基因的检测方法。大多数检测方法都呈现出预期的结果。对低水平检测的能力进行了评估，成功检测到浓度低至 0.06%的元件。本工作中提出的方法将 Fluidigm 系统作为 GMO 多检测的合适且有前途的平台。
