Patent ID: 8224584

Claim:
A system for quantifying an analyte nucleic acid that may be present in a sample undergoing testing, comprising: an apparatus that amplifies nucleic acids and performs time-dependent monitoring of amplicon production, said apparatus comprising a temperature-controlled incubator and an optical detector; a computer operably linked to said apparatus, said computer being programmed to execute the steps of (a) processing results from a test amplification reaction carried out with a test reaction mixture that comprises an internal calibrator and said sample undergoing testing, whereby there is generated a test sample data point comprising first and second coordinates respectively determined by time-dependent indicia of amplification for said analyte nucleic acid and said internal calibrator that coamplified in said test amplification reaction; (b) comparing said test sample data point with a calibration curve to locate a point thereon that comprises first and second coordinates, and that is minimally distant from said test sample data point in a measurement plane that comprises said calibration curve and said test sample data point, wherein said calibration curve comprises (i) first dimension coordinates comprising solutions to a first optimized parametric equation that specifies time-dependent indicia of amplification for an analyte polynucleotide standard as a function of the parameter representing the known starting quantity of said analyte polynucleotide standard in each of a plurality of calibration reactions that comprise said internal calibrator and known starting quantities of said analyte polynucleotide standard, and (ii) second dimension coordinates comprising solutions to a second optimized parametric equation that specifies time-dependent indicia of amplification for said internal calibrator as a function of the parameter representing the known starting quantity of said analyte polynucleotide standard in each of said plurality of calibration reactions, and (c) determining a value for the parameter representing the known starting quantity of said analyte polynucleotide standard that results in said point located on said calibration curve, whereby said value estimates the quantity of said analyte nucleic acid in said sample undergoing testing.