DNA detection method
First Claim
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1. A method for quantitatively detecting ssDNA in a sample, said method comprising:
- combining, in an assay medium, said sample and assay components for detecting said ssDNA, said components comprising;
a high affinity single-tranded DNA binding protein labeled with a detectable label (d1-BP), a high affinity single-stranded DNA binding protein labeled with a specific binding pair member (BP-sbpm) and an immobilized complementary specific binding pair member complementary to said sbpm-BP (s'"'"'bpm), whereby a DNA complex forms between any ssDNA in said sample and said binding proteins, and said BP-sbpm binds to said immobilized s'"'"'bpm; and
detecting immobilized detectable label as indicative of the presence of ssDNA in said sample.
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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.
72 Citations
18 Claims
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1. A method for quantitatively detecting ssDNA in a sample, said method comprising:
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combining, in an assay medium, said sample and assay components for detecting said ssDNA, said components comprising;
a high affinity single-tranded DNA binding protein labeled with a detectable label (d1-BP), a high affinity single-stranded DNA binding protein labeled with a specific binding pair member (BP-sbpm) and an immobilized complementary specific binding pair member complementary to said sbpm-BP (s'"'"'bpm), whereby a DNA complex forms between any ssDNA in said sample and said binding proteins, and said BP-sbpm binds to said immobilized s'"'"'bpm; anddetecting immobilized detectable label as indicative of the presence of ssDNA in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 11, 12, 13, 14)
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8. A method for quantitatively detecting an organism in a sample, said method comprising:
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treating said sample under lysing and denaturing conditions to provide ssDNA in a lysate; contacting said lysate with a positively charged nylon membrane or a nitrocellulose membrane, whereby ssDNA becomes bound to said membrane; contacting said member with a high affinity ssDNA bindign protein conjugated to a detectable label to form complexes with any ssDNA bound to said membrane; and detecting the presence of said detectable label bound to said membrane as indicative of an organism in said sample. - View Dependent Claims (9, 10, 15, 16, 17, 18)
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Specification