Patent US3738365A successfully refreshed from Google Patents.
EP0070685A2
Expired / Lapsed
EP
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Homogeneous nucleic acid hybridization diagnostics by non-radiative energy transfer
Assignee: BP Corp North America Inc
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Filing: 1982-07-14
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Granted: —
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Expires: 2002-07-14
Abstract
Abstract A homogeneous light-emitting hybridization assay is disclosed wherein light-labeled first and second single-stranded reagent segments are hybridized with a complementary target single-stranded polynucleotide from a physiological sample such that non-radiative energy transfer occurs between the light labels of the two reagent segments. At least one of the light labels is of the absorber/emitter type such that energy absorbed from the other light label is re-emitted as a different wavelength. Such secondary emissions can only occur if hybridization has taken place and hence the presence of the target polynucleotide is related to the amount of secondary light emitted.
Inventors
- 👤 Michael James Heller
- 👤 Larry Edward Morrison
- 👤 William Dudley Prevatt
- 👤 Cavit Akin
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 18:42
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6818
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
125 words
A method for assaying a unique polynucleotide sequence or gene segment comprising: a) contacting a single-stranded polynucleotide sample, under hybridization conditions, with first and second single-stranded polynucleotide reagent segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, said first reagent segment having a chemiluminescent catalyst attached thereto and said second reagent segment having an absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide, the chemiluminescent catalyst and the absorber/emitter moiety would be close enough to each other to permit non-radiative energy transfer; b) further contacting the single-stranded polynucleotide sample with chemiluminescent reagents effective for causing light emission in the presence of the chemiluminescent catalyst; and c) measuring the quantity of light emitted by the absorber/emitter moiety.
Claim 2
Independent Claim
125 words
A method for assaying a unique polynucleotide sequence or gene segment comprising: a) contacting a single-stranded polynucleotide sample, under hybridization conditions, with first and second single-stranded polynucleotide reagent segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, said first reagent segment having a chemiluminescent catalyst attached thereto and said second reagent segment having an absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide, the chemiluminescent catalyst and the absorber/emitter moiety would be close enough to each other to permit non-radiative energy transfer; b) further contacting the single-stranded polynucleotide sample with chemiluminescent reagents effective for causing light emission in the presence of the chemiluminescent catalyst; and c) measuring the quantity of light emitted by the absorber/emitter moiety.
Claim 3
Dependent Claim
31 words
The method of Claim 1 wherein the single-stranded polynucleotide sample is contacted with the first and second polynucleotide reagent segments prior to being contacted with the chemiluminescent reagents. or Claim 2
Claim 4
Dependent Claim
22 words
The method of Claim 1 or claim 2 wherein the chemiluminescent catalyst is selected from the group consisting of peroxidase and luciferases.
Claim 5
Dependent Claim
22 words
The method of Claim 1 or Claim 2 wherein the absorber/emitter moiety is selected from the group consisting of fluorophores and phosphores.
Claim 6
Independent Claim
81 words
The method of any preceding claim wherein the first and second single-stranded polynucleotide reagent segments are obtained by: a) cleaving double-stranded target polynucleotides in a physiological sample to form polynucleotide segments; b) denaturing the polynucleotide segments to obtain single-stranded polynucleotide segments; c) end labeling the 3' ends of the polynucleotide segments with either a chemiluminescent catalyst or an absorber/emitter moiety; and d) end labeling the 5' ends of the polynucleotide segments with the catalyst or moiety not used in step (c).
Claim 7
Dependent Claim
35 words
The method of Claim 5 wherein the chemiluminescent catalyst is attached to the 5' ends of the polynucleotide reagent segments and the absorber/emitter moiety is attached to the 3' ends of the polynucleotide reagent segments.
Claim 8
Dependent Claim
32 words
The method of Claim 5 wherein the chemiluminescent catalyst is attached to the 3' ends of the polynucleotide segments and the absorber/emitter moiety is attached to the 5'ends of the polynucleotide segments.
Claim 9
Dependent Claim
91 words
A reagent for detecting the presence of a target single-stranded polynucleotide in a single-stranded polynucleotide sample comprising: a) a first single-stranded polynucleotide reagent segment having at least one chemiluminescent catalyst attached thereto and which is complementary to a portion of a target single-stranded polynucleotide; b) a second single-stranded polynucleotide reagent segment having at least one absorber/emitter moiety attached thereto and which is complementary to a different, substantially mutually exclusive portion of the target single-stranded polynucleotide; and c) chemiluminescent reagents effective for causing light emission in the presence of the chemiluminescent catalyst.
Claim 10
Dependent Claim
91 words
A reagent for detecting the presence of a target single-stranded polynucleotide in a single-stranded polynucleotide sample comprising: a) a first single-stranded polynucleotide reagent segment having at least one chemiluminescent catalyst attached thereto and which is complementary to a portion of a target single-stranded polynucleotide; b) a second single-stranded polynucleotide reagent segment having at least one absorber/emitter moiety attached thereto and which is complementary to a different, substantially mutually exclusive portion of the target single-stranded polynucleotide; and c) chemiluminescent reagents effective for causing light emission in the presence of the chemiluminescent catalyst.
Claim 11
Dependent Claim
21 words
The reagent of Claim 8 wherein a chemiluminescent catalyst is attached at an end of the first single-stranded polynucleotide reagent segment.
Claim 12
Dependent Claim
21 words
The reagent of Claim 8 wherein an absorber/emitter moiety is attached at an end of the second single-stranded polynucleotide reagent segment.
Claim 13
Dependent Claim
20 words
The reagent of Claim 8 wherein the first single-stranded polynucleotide reagent segment contains a chemiluminescent catalyst and an absorber/emitter moiety.
Claim 14
Dependent Claim
23 words
The reagent of Claim 8 wherein both the first and second single-stranded polynucleotide reagent segments contain a chemiluminescent catalyst and an absorber/emitter moiety.
Claim 15
Dependent Claim
27 words
The reagent of Claim 12 wherein the chemiluminescent catalyst and the absorber/emitter moiety are attached to the ends of both the first and second polynucleotide reagent segments.
Claim 16
Dependent Claim
43 words
The reagent of Claim 13 wherein the chemiluminescent catalyst is attached at the 3' end of the first and second single-stranded polynucleotide reagent segments and an absorber/emitter moiety is attached at the 5' end of the first and second single-stranded polynucleotide reagent segments.
Claim 17
Dependent Claim
24 words
The reagent of any on of Claims 8 to 14 wherein the chemiluminescent catalyst is selected from the group consisting of peroxidase and luciferases.
Claim 18
Dependent Claim
24 words
The reagent of any one of Claims 8 to 14 wherein the absorber/emitter moiety is selected from the group consisting of fluorophores and phosphores.
Claim 19
Independent Claim
108 words
A method for assaying a unique polynucleotide sequence or gene segment comprising: a) contacting a single-stranded polynucleotide sample, under hybridization conditions, with a reagent solution comprising first and second single-stranded polynucleotide segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, both of said first and second segments having a different absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide the absorber/emitter moieties would be close enough to each other to permit non-radiative energy transfer; b) irradiating the sample with light sufficient to excite one of the absorber/emitter moieties; and c) measuring the light response from the other absorber/emitter moiety.
Claim 20
Independent Claim
108 words
A method for assaying a unique polynucleotide sequence or gene segment comprising: a) contacting a single-stranded polynucleotide sample, under hybridization conditions, with a reagent solution comprising first and second single-stranded polynucleotide segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, both of said first and second segments having a different absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide the absorber/emitter moieties would be close enough to each other to permit non-radiative energy transfer; b) irradiating the sample with light sufficient to excite one of the absorber/emitter moieties; and c) measuring the light response from the other absorber/emitter moiety.
Claim 21
Dependent Claim
61 words
A reagent solution comprising first and second single-stranded polynucleotide segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, both of said first and second segments having a different absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide the absorber/emitter moieties would be close enough to each other to permit non-radiative energy transfer.
Claim 22
Dependent Claim
61 words
A reagent solution comprising first and second single-stranded polynucleotide segments which are complementary to substantially mutually exclusive portions of a target single-stranded polynucleotide, both of said first and second segments having a different absorber/emitter moiety attached thereto, such that upon hybridization with a target single-stranded polynucleotide the absorber/emitter moieties would be close enough to each other to permit non-radiative energy transfer.
References Cited (Backward Citations)
No citation records indexed.
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