Kit and method for fluorescence based detection assay

• US 6,077,669 A
• Filed: 11/04/1997
• Issued: 06/20/2000
• Est. Priority Date: 11/04/1997
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Little, Michael C., Vonk, Glenn P.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): SITTON, JEHANNE SOUAYA
• Application Number:
  US08/964,020
• Time in Patent Office: 959 Days
• Field of Search: 435/6, 435/91, 435/305.3, 435/968, 435/975, 536/24.3, 536/25.32
• US Class Current: 435/5
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive

Abstract

The present invention relates to kits and methods for conducting fluorescence based detection assays. The kits are configured in a manner to perform the method so as to reduce or eliminate interfering fluorescence signal.

Claims

1. A method for conducting a fluorescence detection assay to determine the presence, absence, or amount of a target analyte in a sample, said method comprising:

(a) producing a mixture by exposing the sample to a first reagent formulation comprising a dry, hydratable binding partner of said target analyte, said binding partner (i) being capable of producing a detectable change of fluorescence signal as the amount of target analyte changes, and (ii) being hydrated by exposure to said sample; and

(b) exposing said mixture to a second reagent formulation or a condition, which, if said target analyte is present in the sample, then the amount of said target analyte will change.

2. The method of claim 1, wherein said target analyte is a nucleic acid molecule.

3. The method of claim 2, wherein said binding partner is a nucleic acid molecule comprising a sequence which hybridizes to the sequence of the target analyte.

4. The method of claim 3 wherein said binding partner comprises a fluorescence energy donor moiety and a fluorescence energy acceptor moiety in sufficiently close proximity that a decrease of such proximity results in the detectable change of fluorescence signal.

5. The method of claim 2, wherein said first reagent formulation further comprises primers for a nucleic acid amplification reaction.

6. The method of claim 5, wherein said binding partner and said primers are in a dry form.

7. The method of claim 2, wherein said second reagent formulation comprises an enzyme necessary for a nucleic acid amplification reaction.

8. The method of claim 7 wherein said enzyme is a polymerase.

9. The method of claim 8 wherein said polymerase is in a dry form.

10. The method of claim 1 wherein said condition is a change of temperature.

11. The method of claim 10 wherein said change of temperature is an increase of temperature.

12. The method of claim 1, further comprising:

(c) containment of said mixture after exposure to said second reagent formulation or said condition.