J. Madic , A. Zocevic , V. Senlis , E. Fradet , B. Andre , S. Muller , R. Dangla , M.E. Droniou
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引用次数: 90
摘要
数字PCR是分子分析的一个令人兴奋的新领域，允许前所未有的核酸定量精度，以及复杂样品中罕见分子事件的精细区分。我们在这里提出了一种新的数字PCR技术，Crystal digital PCR™，它依赖于使用单个芯片将样品划分为二维液滴阵列，然后进行热循环，最后使用三色荧光扫描装置进行读取。因此，这种新技术允许三色复用，这需要一种不同的数据分析方法。在本文中，我们提出了这种创新的工作流程，它既快速又用户友好，并讨论了相关的数据分析问题，如荧光溢出补偿和数据表示。最后，我们还提出了这种三色检测系统的概念验证，使用四重分析来检测EGFR突变L858R, L861Q和T790M。
Digital PCR is an exciting new field for molecular analysis, allowing unprecedented precision in the quantification of nucleic acids, as well as the fine discrimination of rare molecular events in complex samples. We here present a novel technology for digital PCR, Crystal Digital PCR™, which relies on the use of a single chip to partition samples into 2D droplet arrays, which are then subjected to thermal cycling and finally read using a three-color fluorescence scanning device. This novel technology thus allows three-color multiplexing, which entails a different approach to data analysis. In the present publication, we present this innovative workflow, which is both fast and user-friendly, and discuss associated data analysis issue, such as fluorescence spillover compensation and data representation. Lastly, we also present proof-of-concept of this three-color detection system, using a quadriplex assay for the detection of EGFR mutations L858R, L861Q and T790M.
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