Temperature step correction with double sigmoid Levenberg-Marquardt and robust linear regression

摘要

Systems and methods for improving Ct determination in PCR amplification curves by correcting PCR data for temperature shifts that may occur during the PCR process. A double sigmoid function with parameters determined by a Levenberg-Marquardt (LM) regression process is used to find an approximation to the portion of the curve in the region after the temperature shift, termed "CAC", the cycle where the temperature shift occurred. A robust linear approximation is determined for the portion of the curve in the region before the temperature shift. Values of the fluorescent intensity for the cycle CAC or CAC+1 are determined using both the linear approximation and the LM process, and a difference in these values is subtracted off of the portion of the data set representing the portion of the curve before the temperature shift occurred to produce a shift-corrected data set. The shift-corrected data set may be displayed or otherwise used for further processing.