EP0439222A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Water-insoluble reagent, nucleic acid probe, test kit and diagnostic and purification methods
Assignee: Ortho Clinical Diagnostics Inc
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Filing: 1991-01-17
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Granted: —
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Expires: 2011-01-17
Abstract
Abstract An oligonucleotide is linked to a particle through a protein or carbohydrate to form a water-insoluble nucleic acid probe. The protein or carbohydrate has a pI of 6 or less, and has been. chemically modified with an acylating, alkylating or sulfonylating agent. The particle surface is substantially free of other proteins or carbohydrates. The probe is useful in various diagnostic and purification methods wherein hybridization of the oligonucleotide with a target nucleic acid is possible. In one instance, the probe can be used to capture a DNA strand which has been amplified using polymerase chain reaction techniques.
Inventors
- 👤 Harold Chester Iii C/O Eastman Kodak Co. Warren
- 👤 John Bruce C/O Eastman Kodak Co. Findlay
- 👤 Marlene Marie C/O Eastman Kodak Co. King
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-30 12:33
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; C12Q1/6813; C12Q1/6834; C12Q1/70; C12Q1/701; C12Q1/702; C12Q1/703
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
58 words
A water-insoluble reagent comprising a water-insoluble particle having covalently attached thereto a non-immunoreactive protein or carbohydrate, the reagent characterized wherein the non-immunoreactive protein or carbohydrate has a pI of 6 or less, and has been chemically modified with an acylating, alkylating or sulfonylating agent, and the surface of the particle being substantially free of other proteins or carbohydrates.
Claim 2
Dependent Claim
80 words
A water-insoluble nucleic acid probe comprising an oligonucleotide which is attached to a water-insoluble particle, the probe characterized wherein attachment is through a linking group which is a protein or carbohydrate, the protein or carbohydrate having a pI of 6 or less and has been chemically modified with an acylating, alkylating or sulfonylating agent to provide groups for covalent reaction with the oligonucleotide, and is covalently attached to the particle surface which is substantially free of other proteins or carbohydrates.
Claim 3
Dependent Claim
24 words
The probe as claimed in claim 2 wherein the oligonucleotide is a nucleic acid sequence which is complementary with a strand of HIV-I DNA.
Claim 4
Dependent Claim
17 words
The probe as claimed in either of claims 2 and 3 having a detectable label associated therewith.
Claim 5
Dependent Claim
30 words
The invention as claimed in any of claims 1 to 4 wherein the water-insoluble particle is a synthetic polymeric particle having an average diameter of from 0.1 to 10 µmeter.
Claim 6
Dependent Claim
39 words
The invention as claimed in any of claims 1 to 5 wherein the polymeric particles are prepared from monomers having reactive groups selected from the group consisting of carboxy, active halogen atoms, activated 2-substituted ethylsulfonyl, vinylsulfonyl and vinylsulfonylalkylene groups.
Claim 7
Dependent Claim
34 words
The invention as claimed in any of claims 1 to 6 wherein the chemically modified protein or carbohydrate is succinylated casein, glutarylated casein, succinylated collagen, succinylated bovine serum albumin, carboxymethyl cellulose or carboxyethyl cellulose.
Claim 8
Independent Claim
91 words
A method for capturing and isolating a target nucleic acid comprising: A. contacting a test sample suspected of containing a target nucleic acid with the water-insoluble nucleic acid probe as claimed in any of claims 2 to 7, the oligonucleotide of the probe being substantially complementary to the target nucleic acid, the contacting being carried out under hybridization conditions so that hybridization of the target nucleic acid and the oligonucleotide is achieved to form a water-insoluble hybridization product, and B. isolating the hybridized product from the rest of the test sample.
Claim 9
Independent Claim
99 words
A method for the detection of a target nucleic acid comprising: A. contacting a test sample suspected of containing a target nucleic acid with the water-insoluble nucleic acid probe as claimed in any of claims 2 to 7, the oligonucleotide of the probe being substantially complementary to the target nucleic acid, the contacting being carried out under hybridization conditions so that hybridization of the target nucleic acid and the oligonucleotide is achieved to form a water-insoluble hybridization product, and B. detecting the hybridized product as an indication of the presence of the target nucleic acid in the test sample.
Claim 10
Dependent Claim
37 words
The method as claimed in claim 9 carried out as a sandwich hybridization assay wherein the water-insoluble nucleic probe is an unlabeled capture probe, and the target nucleic acid is also hybridized with a water-soluble, labeled probe.
Claim 11
Independent Claim
28 words
The method as claimed in either of claims 9 and 10 wherein the target nucleic acid is amplified using polymerase chain reaction prior to contact in step A.
Claim 12
Dependent Claim
53 words
A diagnostic test kit comprising: (a) the water-insoluble nucleic acid probe as claimed in any of claims 2 to 7, and (b) a water-soluble primer or probe which is substantially complementary to a target nucleic acid, the target nucleic acid also being substantially complementary to the oligonucleotide of the water-insoluble nucleic acid probe.
Claim 13
Dependent Claim
21 words
The test kit as claimed in claim 12 further comprising reagents necessary for polymerase chain reaction of a target nucleic acid.
References Cited (Backward Citations)
No citation records indexed.
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