EP0142299A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Method of measuring polynucleotide and reagent kit for use therein
Assignee: Fujirebio Inc
•
Filing: 1984-10-23
•
Granted: —
•
Expires: 2004-10-23
Abstract
Abstract Disclosed is a method of measuring a polynucleotide such as DNA and RNA in a body fluid which does not require immobilisation of the test sample and whereby the polynucleotide is easily determined, wherein an immobilised singlestranded polynucleotide which is labeled is hybridised with the single-stranded polynucleotide to be measured, the double-stranded polynucleotide formed by hybridisation is broken by means of a restriction enzyme and the labelling substance present is then determined as a measure of the polynucleotide being determined.
Inventors
- 👤 Yoshihiro Ashihara
- 👤 Yasushi Kasahara
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-26 08:03
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6823
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
105 words
A method of measuring a polynucleotide which is either single stranded as such or a single stranded polynucleotide obtained from a double stranded polynucleotide to be measured, which comprises, contacting the single-stranded polynucleotide to be measured with an immobilised labelled single-stranded polynucleotide which is able to hybridise with said single-stranded polynucleotide in aqueous solution to produce a double-stranded polynucleotide, breaking the double-stranded polynucleotide thereby obtained by means of a double-stranded polynucleotide restriction enzyme, and measuring the amount of labelling substance in the solution obtained and/or on immobilised material, as appropriate as a measure of said single stranded and/or said double stranded polynucleotide to be measured.
Claim 2
Independent Claim
105 words
A method of measuring a polynucleotide which is either single stranded as such or a single stranded polynucleotide obtained from a double stranded polynucleotide to be measured, which comprises, contacting the single-stranded polynucleotide to be measured with an immobilised labelled single-stranded polynucleotide which is able to hybridise with said single-stranded polynucleotide in aqueous solution to produce a double-stranded polynucleotide, breaking the double-stranded polynucleotide thereby obtained by means of a double-stranded polynucleotide restriction enzyme, and measuring the amount of labelling substance in the solution obtained and/or on immobilised material, as appropriate as a measure of said single stranded and/or said double stranded polynucleotide to be measured.
Claim 3
Dependent Claim
22 words
The method of claim 1, wherein said single-stranded polynucleotide to be measured is a single-stranded deoxyribonucleic acid or a single-stranded ribonucleic acid.
Claim 4
Dependent Claim
29 words
The method of claim 1 or 2, wherein said labelling substance is selected from radioisotopes, fluorescent materials, luminescent materials, magnetic materials, enzymes, prosthetic groups thereof, coenzymes and enzyme inhibitors.
Claim 5
Dependent Claim
26 words
The method of claim 3, wherein the labelling substance is a large molecule substance and has been introduced into the single-stranded polynucleotide prior to immobilisation thereof.
Claim 6
Independent Claim
35 words
The method of any one of claims 1 to 4, wherein the single-stranded polynucleotide which is labelled has been obtained by denaturation of a double stranded polynucleotide containing the single stranded polynucleotide to be measured.
Claim 7
Independent Claim
37 words
The method of any one of claims 1 to 3, wherein the single stranded polynucleotide which is labelled has been obtained by a solid phase polynucleotide synthesis method or by genetic synthesis with y-DNA using a plasmid.
Claim 8
Independent Claim
30 words
The method of any one of the preceding claims, wherein the carrier material is selected from Sepharose, Sephadex, cellulose gel, ion exchange resin, filter paper, nitrocellulose, nylon, polystyrene and polyacrylamide.
Claim 9
Independent Claim
37 words
The method of any one of the preceding claims, wherein said contacting is carried out at from 20 to 70°C at a pH of from 5 to 9 over a period of from 0.5 to 40 hours.
Claim 10
Independent Claim
22 words
The method of any preceding claim, wherein the restriction enzyme is added to the aqueous solution after hybridisation has been effected therein.
Claim 11
Dependent Claim
43 words
A kit of reagents for use in the method of any preceding claim which comprises an immobilised labelled single stranded polynucleotide which is able to hybridise with a single-stranded polynucleotide in an aqueous solution to produce a double-stranded polynucleotide and a restriction enzyme.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.