US5082935A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Diagnostic reagents made by attaching cytidine containing nucleic acid probes to amino functionalized solid supports by bisulfite mediated transamination
Assignee: BP Corp North America Inc
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Filing: 1988-12-15
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Granted: 1992-01-21
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Expires: 2008-12-15
Abstract
Abstract This invention encompasses diagnostic reagents comprising a polynucleic acid probe having a specific binding sequence and having one or more cytidines outside of the specific binding sequence. The polynucleic acid probe is bound to an amino functionalized solid support by a bisulfite mediated transamination between a cytidine and an amino group on the solid support. When the specific binding sequence contains cytidine, the cytidine is replaced with 5-methylcytidine which has essentially the same binding capacity and yet does not participate in transamination reactions. The invention encompasses compositions of the polynucleic acid probe as well as test kits including the above identified reagent and assays utilizing the above reagent. This invention is useful in detecting viruses, fungi and bacteria in test samples such as body fluids and food samples as well as detecting DNA or RNA sequences in mammalian cells.
Inventors
- 👤 Kenneth A. Cruickshank
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 04:33
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6834; Y; Y10; Y10S; Y10S435/00; Y10S435/808
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
71 words
A diagnostic reagent comprising a polynucleic acid probe having a specific binding sequence and having a cytidine region of one or more cytidines outside of the specific binding sequence and wherein cytidines in the specific binding sequence are 5-methylcytidines, wherein the polynucleic acid probe is bound to an amino functionalized support by a bisulfite mediated transamination between a cytidine outside the specific binding sequence and an amino group on the support.
Claim 2
Dependent Claim
71 words
A diagnostic reagent comprising a polynucleic acid probe having a specific binding sequence and having a cytidine region of one or more cytidines outside of the specific binding sequence and wherein cytidines in the specific binding sequence are 5-methylcytidines, wherein the polynucleic acid probe is bound to an amino functionalized support by a bisulfite mediated transamination between a cytidine outside the specific binding sequence and an amino group on the support.
Claim 3
Dependent Claim
22 words
A diagnostic reagent according to claim 1 wherein the specific binding sequence is complementary to the DNA or RNA to be detected.
Claim 4
Dependent Claim
25 words
A diagnostic reagent according to claim 1 wherein the cytidine region is 1 to 100 cytidines at a terminal end of the polynucleic acid probe.
Claim 5
Dependent Claim
29 words
A diagnostic reagent according to claim 1 wherein the cytidine region is about 5 to 10 cytidines and is not at the terminal end of the polynucleic acid probe.
Claim 6
Dependent Claim
24 words
A diagnostic reagent according to claim 1 wherein the specific binding sequence is a homopolymer sequence selected from polyadenosine, polyguanosine, polythymidine, or poly 5-methylcytidine.
Claim 7
Dependent Claim
18 words
A diagnostic reagent according to claim 1 wherein the specific binding sequence binds to viral DNA or RNA.
Claim 8
Dependent Claim
18 words
A diagnostic reagent according to claim 1 wherein the specific biding sequence binds to bacterial DNA or RNA.
Claim 9
Dependent Claim
18 words
A diagnostic reagent according to claim 1 wherein the specific binding sequence binds to mammalian DNA or RNA.
Claim 10
Dependent Claim
18 words
A diagnostic reagent according to claim 1 wherein the specific binding sequence binds to fungal DNA or RNA.
Claim 11
Dependent Claim
74 words
A diagnostic test kit comprising: (a) a diagnostic reagent of claim 1 wherein the specific binding sequence is complementary to one portion of the DNA or RNA to be determined, or which is complementary to a portion of a polynucleotide bound to a DNA or RNA to be determined and (b) a labeled polynucleic acid probe having a specific binding sequence complementary to a second portion of the DNA or RNA to be determined.
Claim 12
Dependent Claim
36 words
A polynucleic acid probe comprising a specific binding sequence and containing outside of the specific binding sequence a cytidine region of one or more cytidines and wherein the cytidines in the specific binding sequence are 5-methylcytidines.
Claim 13
Dependent Claim
36 words
A polynucleic acid probe comprising a specific binding sequence and containing outside of the specific binding sequence a cytidine region of one or more cytidines and wherein the cytidines in the specific binding sequence are 5-methylcytidines.
Claim 14
Dependent Claim
21 words
A polynucleic acid probe according to claim 11 wherein the specific binding sequence is complementary to the DNA to be determined.
Claim 15
Dependent Claim
22 words
A polynucleic acid probe according to claim 11 wherein the cytidine region is at a terminal end of the polynucleic acid probe.
Claim 16
Dependent Claim
23 words
A polynucleic acid probe according to claim 11 wherein the cytidine region is not at a terminal end of the polynucleic acid probe.
Claim 17
Dependent Claim
17 words
A polynucleic acid probe according to claim 11 wherein the specific binding sequence is a homopolymer sequence.
Claim 18
Dependent Claim
110 words
A method for detecting the presence of specific bacteria in bacteria containing samples comprising: (a) providing a sample suspected of containing the specific bacteria to be detected; (b) lysing the bacterial in said sample to release their DNA; (c) denaturing the DNA; (d) contacting the denatured DNA with a diagnostic reagent of claim 1 and allowing the DNA of the reagent to hybridize with the DNA of the bacteria to be detected; (e) washing the reagent to remove non-specifically bound material; (f) hybridizing a labeled probe to the bacteria DNA bound to the reagent; (g) washing the reagent to remove non-specifically bound DNA; and measuring the label on the reagent.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1995-01-20 | 1995-07-22 | 1996-01-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1999-01-20 | 1999-07-22 | 2000-01-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2003-01-20 | 2003-07-22 | 2004-01-21 | $7700.00 | Window Closed / Lapsed |