Evaluation of uncertainty in quantitative real-time PCR
Evaluation of uncertainty in quantitative real-time PCR
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DOI：
10.1016/j.mimet.2006.04.005
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发表时间：
2006-11-01
影响因子：
2.2
通讯作者：
Samuel, Laly
中科院分区：
文献类型：
--
作者：
Love, John L.;Scholes, Paula;Samuel, Laly
实时定量PCR是用于测量生物系统中的核酸材料的量的较新方法之一。然而，可靠的测量需要一个适当的不确定性估计，本文已经开发了与此程序相关的不确定性预算作为一个例子，从定量实时PCR方法的空肠弯曲菌计数的数据。这种不确定性相对较大，例如，151单位DNA的测量结果将具有+/-84单位DNA的95%置信区间，主要不确定性来源是阈值循环（Ct）值的测量、来自校准线的未知样品的预测DNA含量和DNA的摩尔吸光度值。(c)2006 Elsevier B. V.保留所有权利。
Quantitative real-time PCR is one of the newer methods for measurement of the amount of nucleic material in biological systems. However, reliable measurement requires an appropriate estimation of uncertainty and this paper has developed the uncertainty budget associated with this procedure using as an example, data from a quantitative real-time PCR method for the enumeration of Campylobacter jejuni. This uncertainty is relatively large and for instance, a measured result of 151 units of DNA would have a 95% confidence interval of +/- 84 units of DNA with the main sources of uncertainty being the measurement of the threshold cycle (Ct) value, the predicted DNA content of the unknown sample from the calibration line and the molar absorbance value for DNA. (c) 2006 Elsevier B.V. All rights reserved.