US 5,283,174 AGranted

Homogenous protection assay

AssigneeGen-Probe Incorporated

InventorsLyle J. Arnold, Jr., Norman C. Nelson

Life of this patent

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Abstract

Improved homogenous diagnostic assay methods and labels for detecting an analyte in a medium when the analyte is a member of a specific binding pair. The methods and labels provide procedures for reducing background and increasing sensitivity. The binding partner of the analyte is labeled with a substance, the stability of which detectably changes whenever said analyte is bound as a member of the specific binding pair. In a closely related system, the analyte is labeled with a substance susceptible to differential degradation depending on whether or not the analyte is bound as a member of its specific binding pair. After incubation and selective degradation or chemical or biochemical alteration, the amount of analyte bound is detected by measuring either the stability change or the extent of degradation of the label. In a particular system, chemiluminescent acridinium ester labeled probes are used in a homogenous hybridization assay format for sensitively detecting the presence of complement any target polynucleotide sequences.

Claims12

Dependent claims

A homogeneous assay method for determining the presence or amount of a first nucleic acid in a medium, comprising the steps of:

contacting a free second nucleic acid with said medium to form a hybridization mixture, wherein said free second nucleic acid is able to hybridize with said first nucleic acid under hybridizing conditions to form a binding pair, and wherein said second nucleic acid is chemically bonded to an N-acridinium ester which is selectively susceptible to chemical degradation by a chemical selected from the group consisting of an acid, a base, and an oxidizing agent, to form an altered N-acridinium ester dependent upon whether said second nucleic acid is free or forms part of said binding pair;
treating said hybridization mixture with said chemical such that N-acridinium ester chemically bonded to free second nucleic acid is degraded to form altered N-acridinium ester and N-acridinium ester present in a binding pair is undegraded;
and
determining the amount of undegraded N-acridinium ester remaining after said treating step, without physically separating any said free second nucleic acid from any said binding pair, as a measure of the presence or amount of said first nucleic acid.

1 more element

Dependencies are inferred from the claim text.

Facts

Application: 07/613,603Filed 8 Nov 1990 · continuation
Priority: 21 Sep 1987
Granted: 1 Feb 1994
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

G01N 33/582
Involving labelled substances: with fluorescent label
C12Q 1/6816
Hybridisation assays: characterised by the detection means
C12Q 1/6886
For diseases caused by alterations of genetic material: for cancer
C12Q 1/689
For detection or identification of organisms: for bacteria

Family

Serial99,392Parent of this continuation-in-part · filed 21 Sep 1987
Serial99,392Parent of this continuation · filed 21 Sep 1987
Serial294,700Parent of this continuation · filed 12 Dec 1988
Serial528,920Parent of this continuation · filed 23 May 1990
US 5,283,174 AThis patent

From the US filing history. Foreign family members aren’t listed.