Signal enhancement method and kit

• US 6,110,682 A
• Filed: 11/30/1998
• Issued: 08/29/2000
• Est. Priority Date: 10/21/1996
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Dellinger, Douglas J., Dahm, SueAnn C., Troll, Mark A.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US09/201,674
• Time in Patent Office: 638 Days
• Field of Search: 435/6, 536/23.1, 536/24.3, 935/76, 935/77, 935/78
• US Class Current: 435/6.19
• CPC Class Codes: C12Q 1/682 Signal amplification
• CPC Class Codes: C12Q 2525/179 incorporating arbitrary or random nucleotide sequences
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2565/102 Multiple non-interacting labels

Abstract

The invention discloses and claims a signal amplification method for detecting a target nucleic acid analyte having a homopolymeric region and a target sequence. The method comprises (a) contacting an analyte under hybridizing conditions with a multiplicity of reporter probes, each probe including a signal region and an oligonucleotide sequence which is complementary to, and capable of forming a stable hybrid with the analyte homopolymeric region; and (b) forming an analyte:capture probe hybrid by contacting the analyte target sequence with a capture probe under hybridizing conditions.

Claims

1. A method comprising the steps of:

(a) contacting a target nucleic acid analyte having a homopolymeric region and a target sequence, under hybridizing conditions with a multiplicity of reporter probes, said probes comprising a signal region and an oligonucleotide sequence, wherein said oligonucleotide sequence is complementary to said analyte homopolymeric region, so as to form a stable hybrid of multiple reporter probes with said analyte homopolymeric region; and

(b) forming an analyte;

capture probe hybrid by contacting said analyte target sequence with a capture probe under hybridizing conditions.

2. The method of claim 1, wherein said reporter probes hybridize with said analyte homopolymeric region to form a double helix.

3. The method of claim 1, wherein said reporter probe comprises a fluorescent polymer.

4. The method of claim 3, wherein said fluorescent polymer is selected from the group consisting of protein, oligonucleotide and polysaccharide.

5. The method of claim 4, wherein said fluorescent polymer comprises a fluorophore, an oligonucleotide and a linker for joining said oligonucleotide and said fluorophore.

6. The method of claim 5, wherein:

(a) said fluorescent oligonucleotide is a linear polymer comprised of (dT)_n, (U)_n or (U'"'"')_n, where n is an integer between 8 and 30; and

(b) the 3'"'"' terminus of said target nucleic acid analyte has a homopolymeric region comprised of poly A.

7. The method of claim 4, wherein said polymer is a fluorescent protein.

8. The method of claim 7, wherein said fluorescent protein is phycoerythrin.