Patent ID: 8275556
Filing Date: 2012-09-25
Classification: C12Q,G16B,Y10T

Abstract:
1. A method for quantifying an analyte polynucleotide in a test sample by internal calibration adjustment, comprising the steps of: (a) obtaining at least coefficients of an equation for a first calibration curve prepared using a first apparatus that amplifies nucleic acids and monitors amplicon production as amplification is occurring to yield time-dependent indicia of amplification, (b) obtaining a first adjustment calibrator that comprises a first known quantity of the analyte polynucleotide standard and said constant starting quantity of the internal calibrator; (c) coamplifying the internal calibrator and the analyte polynucleotide standard of the first adjustment calibrator in an in vitro nucleic acid amplification reaction performed on a second apparatus that amplifies nucleic acids and monitors amplicon production as amplification is occurring, (d) calculating the ratio of time-dependent indicia of amplification for the analyte polynucleotide standard to time-dependent indicia of amplification for the internal calibrator determined in step (c); (e) calculating a first adjustment factor as the difference between the ratio calculated in step (d), and (f) establishing an adjusted equation for a second calibration curve by modifying the baseline adjustment coefficient of the equation for the first calibration curve using the first adjustment factor calculated in step (e); (g) coamplifying the analyte polynucleotide of the test sample and said constant starting quantity of the internal calibrator in a test reaction performed on the second apparatus to determine time-dependent indicia of amplification for the analyte polynucleotide and the internal calibrator of the test reaction; (h) calculating the ratio of time-dependent indicia of amplification for the analyte polynucleotide to time-dependent indicia of amplification for the internal calibrator determined in the test reaction; and (i) comparing the ratio calculated in step (h) with the second calibration curve, thereby quantifying the analyte polynucleotide in the test sample.