DNA detection method

• US 4,963,658 A
• Filed: 12/15/1988
• Issued: 10/16/1990
• Est. Priority Date: 09/04/1987
• Status: Expired due to Term

• Current Assignee: Molecular Devices
• Original Assignee: Molecular Devices
• Inventors: Kung, Viola T., Sheldon, Edward L. III, Nagainis, Peter A.
• Primary Examiner(s): Nucker, Christine
• Assistant Examiner(s): Krupen, Karen I.
• Application Number:
  US07/285,895
• Time in Patent Office: 670 Days
• Field of Search: 530/402, 530/406, 530/387, 530/350, 435/6, 435/7, 435/177, 436/501
• US Class Current: 435/233
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins
• CPC Class Codes: C12Q 2563/125 the label being enzymatic, i.e. proteins, and non proteins, such as nucleic acid with enzymatic activity
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Abstract

Picogram amounts of DNA can be detected in a sample by the use of high-affinity, single-stranded DNA binding proteins. The assay is applicable not only to pure DNA samples but also to samples containing significant amounts of protein.

Claims

1. A conjugate comprising a high affinity single-stranded DNA binding protein (BP) covalently linked to a detectable label (dl), wherein said conjugate is capable of binding with high affinity via said BP directly to DNA to be detected.

2. The conjugate according to claim 1, wherein said BP is single-stranded DNA binding protein, T4 gene 32 protein, or topoisomerase I.

3. The conjugate according to claim 2, wherein said enzyme is urease or horseradish peroxidase.

4. The conjugate according to claim 1, wherein said dl is an enzyme.

5. A conjugate comprising a high affinity single-stranded DNA binding protein (BP) covalently linked to a specific binding pair member (sbpm), wherein said conjugate is capable of binding with high affinity via said BP directly to DNA to be detected.

6. The conjugate according to claim 5, wherein said sbpm is biotin, avidin, streptavidin or antibiotin.