Enhanced analysis of real-time PCR data by using a variable efficiency model: FPK-PCR.
Enhanced analysis of real-time PCR data by using a variable efficiency model: FPK-PCR.
复制标题
DOI：
10.1093/nar/gkr775
复制
发表时间：
2012-01
影响因子：
14.9
通讯作者：
Goetghebeur E
中科院分区：
文献类型：
--
作者：
Lievens A;Van Aelst S;Van den Bulcke M;Goetghebeur E
目前的实时聚合酶链反应（PCR）分析方法表现良好，只要PCR效率在反应过程中保持恒定。然而，效率的微小变化可能导致大的量化误差。特别是在生物样品中，抑制剂的可能存在形成了挑战。本文提出了一种新的单反应效率计算方法，称为全过程动力学PCR（FPK-PCR）。它结合了一个动力学更现实的模型与灵活的适应全范围的数据。通过重建整个循环效率链，而不是将重点限制在“应用窗口”上，人们可以提取额外的信息，并失去一定程度的任意性。该模型返回的最大效率估计值在准确度和精密度方面与系列稀释的金标准和其他单一反应效率方法相当。如FPK-PCR程序所描述的，循环间效率的变化比其他S形模型的数据更接近数据。单个循环效率的评估比其他单一效率方法返回更多的信息。它可以深入解读实时PCR数据并重建荧光数据，从而提供质量控制。最后，通过实施全局效率模型，可再现性得到改善，因为避免了应用窗口的选择。
Current methodology in real-time Polymerase chain reaction (PCR) analysis performs well provided PCR efficiency remains constant over reactions. Yet, small changes in efficiency can lead to large quantification errors. Particularly in biological samples, the possible presence of inhibitors forms a challenge. We present a new approach to single reaction efficiency calculation, called Full Process Kinetics-PCR (FPK-PCR). It combines a kinetically more realistic model with flexible adaptation to the full range of data. By reconstructing the entire chain of cycle efficiencies, rather than restricting the focus on a ‘window of application’, one extracts additional information and loses a level of arbitrariness. The maximal efficiency estimates returned by the model are comparable in accuracy and precision to both the golden standard of serial dilution and other single reaction efficiency methods. The cycle-to-cycle changes in efficiency, as described by the FPK-PCR procedure, stay considerably closer to the data than those from other S-shaped models. The assessment of individual cycle efficiencies returns more information than other single efficiency methods. It allows in-depth interpretation of real-time PCR data and reconstruction of the fluorescence data, providing quality control. Finally, by implementing a global efficiency model, reproducibility is improved as the selection of a window of application is avoided.
登录
查看更多内容
影响因子：
1.6
作者：
Opel, Kerry L.;Chung, Denise;McCord, Bruce R.
通讯作者：
McCord, Bruce R.
影响因子：
3
作者：
Alvarez MJ;Vila-Ortiz GJ;Salibe MC;Podhajcer OL;Pitossi FJ
通讯作者：
Pitossi FJ
影响因子：
3.8
作者：
Gevertz, JL;Dunn, SM;Roth, CM
通讯作者：
Roth, CM
影响因子：
3.3
作者：
Barbau-Piednoir, Elodie;Lievens, Antoon;Van den Bulcke, Marc
通讯作者：
Van den Bulcke, Marc
影响因子：
2
作者：
Jagers, P;Klebaner, F
通讯作者：
Klebaner, F