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PCR elbow determination by use of a double sigmoid function curve fit with the Levenberg-Marquardt algorithm and normalization

机译:通过使用符合Levenberg-Marquardt算法的双S形函数曲线确定PCR肘并进行归一化

摘要

Systems and methods for determining characteristic transition values such as elbow values in sigmoid or growth-type curves, such as the cycle threshold (Ct) value in PCR amplification curves. A double sigmoid function with parameters determined by a Levenberg-Marquardt (LM) regression process is used to find an approximation to a curve that fits a PCR dataset. Once the parameters have been determined, the curve can be normalized using one or more of the determined parameters. Normalization is advantageous for determining the Ct value if one chooses the arbitrary fluorescence level (AFL) approach to calculating Ct values for amplification curves. After normalization, the normalized curve is processed by applying a root-finding algorithm to determine the root of the function representing the normalized curve, which root corresponds to the Ct value. The Ct value is then returned and may be displayed or otherwise used for further processing.
机译:用于确定特征性过渡值(例如S型曲线中的弯头值或生长型曲线)(例如PCR扩增曲线中的循环阈值(Ct)值)的系统和方法。使用由Levenberg-Marquardt(LM)回归过程确定的参数的双S型函数来查找拟合PCR数据集的曲线的近似值。一旦确定了参数,就可以使用一个或多个确定的参数对曲线进行归一化。如果选择任意荧光水平(AFL)方法来计算扩增曲线的Ct值,则标准化对于确定Ct值是有利的。归一化之后,通过应用根查找算法来处理归一化曲线,以确定代表归一化曲线的函数的根,该根对应于Ct值。然后返回Ct值,可以显示Ct值或将其用于其他处理。

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