Measuring Digital PCR Quality: Performance Parameters and Their Optimization
Measuring Digital PCR Quality: Performance Parameters and Their Optimization
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DOI：
10.1371/journal.pone.0153317
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发表时间：
2016-05-05
期刊：
影响因子：
3.7
通讯作者：
Querci, M.
中科院分区：
文献类型：
--
作者：
Lievens, A.;Jacchia, S.;Querci, M.
数字PCR在基于DNA的食品分析领域正迅速被采用。它提供的直接、绝对定量使其成为一种有吸引力的技术，用于食品和饲料样品的常规分析，以确定其成分、可能的转基因成分和是否符合标签要求。然而，尚未充分确立dPCR检测试剂盒性能的评估。本文介绍了基于统计学原理的三个简单参数，允许用户评估其分析是否稳健。此外，我们提出了运行后评价标准，以检查定量是否准确。最后，我们评估了泊松置信区间的有用性，并提出了一种替代策略，以更好地捕捉分析链中的变异性。
Digital PCR is rapidly being adopted in the field of DNA-based food analysis. The direct, absolute quantification it offers makes it an attractive technology for routine analysis of food and feed samples for their composition, possible GMO content, and compliance with labelling requirements. However, assessing the performance of dPCR assays is not yet well established. This article introduces three straightforward parameters based on statistical principles that allow users to evaluate if their assays are robust. In addition, we present post-run evaluation criteria to check if quantification was accurate. Finally, we evaluate the usefulness of Poisson confidence intervals and present an alternative strategy to better capture the variability in the analytical chain.