Nucleic acid detection with energy transfer

{
  "publication_number": "US5827653A",
  "country": "US",
  "kind": "A",
  "title": "Nucleic acid detection with energy transfer",
  "abstract": "PCT No. PCT/GB94/02068 Sec. 371 Date May 20, 1996 Sec. 102(e) Date May 20, 1996 PCT Filed Sep. 23, 1994 PCT Pub. No. WO95/08642 PCT Pub. Date Mar. 30, 1995A method for the detection of a nucleic acid analyte by complementary probe hybridisation and formation of a chelated lanthanide complex which, upon irradiation by light, results in a characteristic delayed luminescence emission.",
  "claims": [
    "1. A method for detecting a nucleic acid analyte in a sample, which method comprises contacting the sample with (i) a binding entity complementary to the analyte and to which is attached a first partner of a ligand to metal energy transfer system, (ii) a lanthanide ion, and (iii) a second partner of the system to which is attached a duplex binder; wherein the first partner of the ligand to metal energy transfer system is a complexing sensitiser and the second partner is a chelating agent, or vice versa; such that upon complementary binding of the analyte and the binding entity to form a nucleic acid duplex, interaction of the duplex binder with the duplex occurs and allows the first and the second partners of the system to form a closed chelated system around the lanthanide ion and, upon irradiation, emission of light from the lanthanide ion indicates the presence of the analyte in the sample.",
    "2. A method as claimed in claim 1 wherein the duplex binder is selected from the group consisting of an intercalator and a groove binder.",
    "3. A method as claimed in claim 1 wherein the binding entity is a polynucleotide to which the first or the second partner is attached.",
    "4. A method as claimed in claim 3 wherein the binding entity comprises a blocked terminal nucleotide.",
    "5. A method as claimed in any one of the previous claims wherein the binding entity comprises one or more mismatched bases opposite a complementary analyte nucleic acid sequence.",
    "6. A method as claimed in claim 5 wherein a position of the one or more mismatched bases is selected such that duplex binding and hence formation of the closed chelated system around the lanthanide ion are modulated.",
    "7. A method as claimed in claim 1 wherein the lanthanide ion is an europium ion.",
    "8. A method as claimed in claim 1 wherein the complexing sensitiser comprises an aromatic group.",
    "9. A method as claimed in claim 1 wherein the complexing sensitiser comprises a heterocyclic aromatic group.",
    "10. A method as claimed in claim 1 wherein the complexing sensitiser comprises an optionally substituted dipyridyl or phenanthroline group.",
    "11. A method as claimed in claim 1 wherein the complexing sensitiser comprises an optionally substituted phenanthroline-2,6-dicarboxylic acid group.",
    "12. A method as claimed in claim 1 wherein the chelating agent is chelated with the lanthanide ion before addition to the assay medium.",
    "13. A method as claimed in claim 1 wherein the binding entity is a polynucleotide to which a chelated lanthanide ion is attached.",
    "14. A method as claimed in claim 1 wherein one partner of the ligand to metal energy transfer system is a complexing sensitiser linked to a duplex binder.",
    "15. A method as claimed in claim 1 wherein the binding entity is an allele-specific polynucleotide.",
    "16. A method as claimed in claim 1 which comprises amplification of a target sequence to provide the nucleic acid analyte.",
    "17. A method as claimed in claim 16 wherein one or more components of the method are present during amplification of the target sequence.",
    "18. A method as claimed in claim 1 wherein the binding entity is comprised of two or more polynucleotide sequences.",
    "19. A method as claimed in claim 18 wherein the first and the second partners of the ligand to metal energy transfer system are attached to different polynucleotide sequences.",
    "20. A method as claimed in claim 1 wherein the chelating agent is a polyfunctional compound comprising carboxylic acid, amide and/or ether moieties.",
    "21. A method as claimed in claim 20 wherein the chelating agent is a derivative of a compound selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA) and trans-1,2-diaminocyclohexanetetraacetic acid (DCTA).",
    "22. A method as claimed in claim 1 wherein a Forster energy transfer acceptor is used to receive energy from the closed chelated system around the lanthanide ion and to emit light.",
    "23. A method as claimed in claim 22 wherein the Forster energy transfer acceptor is bound to a polynucleotide.",
    "24. A method as claimed in claim 1 for detection of more than one nucleic acid analyte in a sample.",
    "25. A method as claimed in claim 1 wherein the characteristics of light emission are measured to indicate the nature and/or quantity of the nucleic acid analyte.",
    "26. A method as claimed in claim 1 which is a homogeneous assay.",
    "27. An assay kit which comprises (i) a binding entity complementary to the analyte and to which is attached a first partner of a ligand to metal energy transfer system, (ii) a lanthanide ion, (iii) a second partner of the system to which is attached a duplex binder; wherein the first partner of the ligand to metal energy transfer system is a complexing sensitiser and the second partner is a chelating agent, or vice versa; and further comprising one or more of the following: buffer(s), amplification primers and instructions for use of the kit.",
    "28. An assay kit as claimed in claim 27 wherein the amplification primers are for allele specific amplification.",
    "29. A compound which comprises a duplex binder covalently linked to a lanthanide ion sensitiser.",
    "30. A compound as claimed in claim 29 which comprises a phenanthroline-2,6-dicarboxylic acid group linked to a phenanthridine group.",
    "31. A compound as claimed in claim 30 of the formula ##STR3##",
    "32. A composition which comprises a duplex binder covalently linked to a lanthanide ion chelating agent and a lanthanide ion.",
    "33. A compound as claimed in any one of claims 27 and 29-31 wherein the duplex binder is selected from the group consisting of an intercalator and a groove binder."
  ],
  "cpc": [
    "C07D 471/04",
    "C07H 21/00",
    "C12Q 1/6818",
    "G01N 2458/40",
    "Y10S 435/81",
    "Y10S 436/805"
  ],
  "assignees": [
    "ZENECA LTD"
  ],
  "filing_date": "1994-09-23",
  "publication_date": "1998-10-27",
  "grant_date": "1998-10-27",
  "priority_date": "1993-09-23",
  "application_number": "US-61972496-A",
  "family_id": "26303577",
  "citations": [
    "DE4119075A1",
    "EP0144914A2",
    "EP0171978A1",
    "EP0242527A2",
    "US4665184A",
    "US4942227A",
    "US4954714A",
    "US5565554A",
    "US5571897A",
    "US5578498A",
    "WO8904375A1",
    "WO9216840A1"
  ]
}