Method of compensation of dose-response curve of an essay for sensitivity to perturbing variables

摘要

The present invention provides assays and methods of compensating for changes in the dose-response curve of an assay where such changes are due to variations in a perturbing variable such as, but not limited to temperature. This is achieved by a two-step method, the first step of which involves measurements of the dose-response curve, and thus the individual assay parameters, at many different values of the perturbing variable, spanning the expected range of the perturbing variable. In the second step, unknown samples are assayed simultaneously with a known standard at a chosen analyte concentration. During this measurement, the value of the perturbing variable is unknown and the dose-response curve is therefore also unknown. The different dose-response curves from the first step are used to determine a mathematical relationship between the assay parameters and the assay signal of the known standard. With this relationship, in which the value of the perturbing variable is implicit rather than explicit, assay parameters that are valid for the unknown value of the perturbing variable can be obtained by substituting the value of the assay signal from the known standard (measured when assaying the unknown samples) into the mathematical relationship and solving for the assay parameters. The method enables an accurate determination of the analyte concentration even when the perturbing variable is changing or fluctuating from one sample measurement to another. Once the first step is completed, the second step can be performed repeatedly to measure unknown samples with accuracy.