US 5,206,179 A · Fluorescence polarization immunoassays for multiple analytes using sequential addition of tracers

Abstract

A methodology is presented which relates in general to fluorescence polarization immunoassays (FPIA) and modifications of current processes wherein detection and quantification of several commonly abused or therapeutic drugs in a single biological fluid sample are determined utilizing manual or current software or related equipment to allow the sequential and simultaneous performance of more than one FPIA assay. The methodology involves combining the reagents either separately or pre-mixed, for multiple assays in a single reagent package, these reagents being used to assay quantitative amounts of each of the assay analytes in a sequential step manner. The assay being performed by mixing the sample with a combination reagent and then initiating a specific reaction for each of the separate analytes by sequentially adding a specific reagent, i.e. tracer for each and reading the results during each separate stage to determine the specific reaction taken place and utilization of manual or software modification monitoring to subtract out the contribution to total signal of each specific reagent added previously to the stage of interest.

Claims11

Dependent claims

A method for performing multiple fluorescence polarization immunoassays of a biological fluid sample contained in a single container wherein the presence of specific analytes (A.sub.n) where n is an integer of 2 or greater, and their respective quantitative amounts are determined, comprising:

(a) contacting the sample and a combination of at least two antiserum reagents (S.sub.n wherein n is an integer of 2 or greater) from at least two or more assays, incubating the resultant mixture of A.sub.n and S.sub.n to an equilibrium state;
(b) taking a fluorescence reading, recording said reading as a blank reading B.sub.1 ;
(c) adding a tracer T.sub.1 and incubating the mixture of A.sub.n, S.sub.n and T.sub.1 to an equilibrium state;
(d) taking a fluorescence reading R.sub.1, recording said reading R.sub.1 as B.sub.2 ;
(e) adjusting the R.sub.1 reading by subtraction of B.sub.1 and calculating a mP value;
and
(f) repeating at least once steps (c), (d) and (e) using T.sub.2, B.sub.2 and sequentially T.sub.2+ m and B.sub.2+m wherein m is an integer of 1 or greater.

4 more elements

(a) contacting the sample and a combination of at least two antiserum reagents (S.sub.n wherein n is an integer of 2 or greater) from at least two or more assays, incubating the resultant mixture of A.sub.n and S.sub.n to an equilibrium state;
(b) taking a fluorescence reading, recording said reading as a blank reading B.sub.1 ;
(c) adding a tracer T.sub.1 and incubating the mixture of A.sub.n, S.sub.n and T.sub.1 to an equilibrium state;
(d) taking a fluorescence reading R.sub.1, recording said reading R.sub.1 as B.sub.2 ;
(e) adjusting the R.sub.1 reading by subtraction of B.sub.1 and calculating a mP value;
and
(f) repeating at least once steps (c), (d) and (e) using T.sub.2, B.sub.2 and sequentially T.sub.2+m and B.sub.2+m wherein m is an integer of 1 or greater;
and
(g) maintaining concentration of each antiserum reagent S.sub.n and tracer T.sub.n wherein n is an integer of one or greater by addition of S.sub.n and T.sub.n after each mP calculation.

6 more elements

Dependencies are inferred from the claim text.

Fluorescence polarization immunoassays for multiple analytes using sequential addition of tracers

Life of this patent

Fluorescence polarization immunoassays for multiple analytes using sequential addition of tracers

Life of this patent

Abstract

Claims11