US4751177A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Methods and kits for performing nucleic acid hybridization assays
Assignee: Amgen Inc
•
Filing: 1985-06-13
•
Granted: 1988-06-14
•
Expires: 2005-06-13
Abstract
Abstract A method for the isolation and quantitative detection of a selected single-stranded target polynucleotide from solution. The target polynucleotide is hybridized in solution to a single-stranded mediator polynucleotide, a probe polynucleotide, and an immobilized polynucleotide sequence. The sequence of the mediator polynucleotide comprises a first sequence complementary to a first portion of the target polynucleotide sequence and a second nucleotide sequence complementary to a portion of a single-stranded immobilized polynucleotide sequence. The probe polynucleotide, which carries a detectable label, is complementary to a second portion of the necleotide sequence of the target. The immobilized polynucleotide is immobilized by attachment to a solid support, and, through hybridization to the mediator polynucleotide, functions to immobilize the entire immobilized polynucleotide/target polynucleotide/probe polynucleotide "sandwich".
Inventors
- 👤 Yitzhak Stabinsky
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 01:39
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C12Q1/705; C12Q1/68; C12Q1/6813; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/814; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/824; Y10S930/00; Y10S930/01; Y10S930/22; Y10S930/224
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
169 words
A method for searching a sample solution for selected target polynucleotide suspended in the solution comprising the steps of: (a) exposing the sample to a mediator polynucleotide having a first portion with a nucleotide sequence complementary to and hybridizable with an immobilized polynucleotide and having a second portion, separate and distinct from the first portion, with a nucleotide sequence complementary to and hybridizable with a first portion of the target polynucleotide; (b) introducing to the sample a probe polynucleotide having an attached reporter group and having a portion with a nuceotide sequence complementary to and hybridizable with a second portion of the target polynucleotide, said second portion being separate and distinct from said first portion of the target polynucleotide; (c) hybridizing the first portion of the target with the second portion ofthe mediator polynucleotide and the second portion of the target polynucleotide with the portion of the probe polynucleotide; and (d) immobilizing the mediator polynucleotide by hybridization of the first portion of the mediator polynucleotide with the immobilized polynucleotide.
Claim 2
Independent Claim
169 words
A method for searching a sample solution for selected target polynucleotide suspended in the solution comprising the steps of: (a) exposing the sample to a mediator polynucleotide having a first portion with a nucleotide sequence complementary to and hybridizable with an immobilized polynucleotide and having a second portion, separate and distinct from the first portion, with a nucleotide sequence complementary to and hybridizable with a first portion of the target polynucleotide; (b) introducing to the sample a probe polynucleotide having an attached reporter group and having a portion with a nuceotide sequence complementary to and hybridizable with a second portion of the target polynucleotide, said second portion being separate and distinct from said first portion of the target polynucleotide; (c) hybridizing the first portion of the target with the second portion ofthe mediator polynucleotide and the second portion of the target polynucleotide with the portion of the probe polynucleotide; and (d) immobilizing the mediator polynucleotide by hybridization of the first portion of the mediator polynucleotide with the immobilized polynucleotide.
Claim 3
Dependent Claim
27 words
The method as recited in claim 1 further comprising the step of qualitating the amount of signal supplied by said detectable label on said immobilized hybrid product.
Claim 4
Dependent Claim
14 words
The method as recited in claim 1 wherein said target polynucleotide is a polydeoxyribonucleotide.
Claim 5
Dependent Claim
14 words
The method as recited in claim 1 wherein said target polynucleotide is a polyribonucleotide.
Claim 6
Dependent Claim
19 words
The method as recited in claim 3 or 4 wherein said mediator polynucleotide sequence is a deoxyribonucleic acid sequence.
Claim 7
Dependent Claim
18 words
The method as recited in claim 3 or 4 wherein said mediator polynucleotide sequence is ribonucleic acid sequence.
Claim 8
Dependent Claim
19 words
The method as recited in claim 3 or 4 wherein said probe polynucleotide sequence is a deoxyribonucleic acid sequence.
Claim 9
Dependent Claim
19 words
The method as recited in claim 3 or 4 wherein said probe polynucleotide sequence is a ribonucleic acid sequence.
Claim 10
Dependent Claim
16 words
The method as recited in claim 3 or 4 wherein said immobilized polynucleotide is a polydeoxyribonucleotide.
Claim 11
Dependent Claim
16 words
The method as recited in claim 3 or 4 wherein said immobilized nucleotide is a polyribonucleotide.
Claim 12
Dependent Claim
138 words
A kit for performing a hybridization assay on a sample to search for a selected target polynucleotide comprising: a polynucleotide immobilized on a support; a mediator polynucleotide, associated as a component of a kit with said immobilized polynucleotide, having a first portion with a nucleotide sequence complementary to and hybridizable with said immobilized polynucleotide and having a second portion, separate and distinct from the first portion, with a nucleotide sequence complementary to and hybridizable with a first portion of the target polynucleotide; a polynucleotide probe, associated as a component of a kit with said immobilized polynucletide, having a portion with a nucleotide sequence complementary to and hybridizable with a second portion of the target polynucleotide, said second portion being separate and distinct from said first portion of the target polynucleotide; and a reporter attached to said probe polynucleotide.
Claim 13
Dependent Claim
138 words
A kit for performing a hybridization assay on a sample to search for a selected target polynucleotide comprising: a polynucleotide immobilized on a support; a mediator polynucleotide, associated as a component of a kit with said immobilized polynucleotide, having a first portion with a nucleotide sequence complementary to and hybridizable with said immobilized polynucleotide and having a second portion, separate and distinct from the first portion, with a nucleotide sequence complementary to and hybridizable with a first portion of the target polynucleotide; a polynucleotide probe, associated as a component of a kit with said immobilized polynucletide, having a portion with a nucleotide sequence complementary to and hybridizable with a second portion of the target polynucleotide, said second portion being separate and distinct from said first portion of the target polynucleotide; and a reporter attached to said probe polynucleotide.
Claim 14
Dependent Claim
14 words
The kit as recited in claim 11 wherein said target polynucleotide is a polydeoxyribonucleotide.
Claim 15
Dependent Claim
14 words
The kit as recited in claim 11 wherein said target polynucleotide is a polyribonucleotide.
Claim 16
Dependent Claim
19 words
The kit as recited in claim 12 or 13 wherein said mediator polynucleotide sequence is a deoxyribonucleic acid sequence.
Claim 17
Dependent Claim
19 words
The kit as recited in claim 12 or 13 wherein said mediator polynucleotide sequence is a ribonucleic acid sequence.
Claim 18
Dependent Claim
19 words
The kit as recited in claim 12 or 13 wherein said probe polynucleotide sequence is a deoxyribonucleic acid sequence.
Claim 19
Dependent Claim
19 words
The kit as recited in claim 12 or 13 wherein said probe polynucleotide sequence is a ribonucleic acid sequence.
Claim 20
Dependent Claim
16 words
The kit as recited in claim 12 or 13 wherein said immobilized polynucleotide is a polydeoxyribonucleotide.
Claim 21
Dependent Claim
16 words
The kit as recited in claim 12 or 13 wherein said immobilized polynucleotide is a polyribonucleotide.
References Cited (Backward Citations)
US4358535A
Cited
US4358535B1
Cited
US4415732A
Cited
EP0062286A1
Cited
US4486539A
Cited
US4522922A
Cited
WO1984003520A1
Cited
EP0130515A2
Cited
EP0153873A2
Cited
WO1985004674A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-06-14 | 1991-12-14 | 1992-06-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-06-14 | 1995-12-14 | 1996-06-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-06-14 | 1999-12-14 | 2000-06-14 | $7700.00 | Window Closed / Lapsed |