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Methods and compositions for universal detection of nucleic acids

  • US 20110212846A1
  • Filed: 02/09/2011
  • Published: 09/01/2011
  • Est. Priority Date: 02/09/2010
  • Status: Active Grant
First Claim
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1. A method of detecting the presence or amount of a first target nucleic acid in a sample, the method comprising:

  • (a) linking first and second universal DNA segments into a first molecule in a linking reaction dependent on the first target nucleic acid, thereby providing a first tagged target nucleic acid; and

    (b) PCR amplifying the first tagged target nucleic acid using first and second universal primers, each universal primer having a 3′

    portion that anneals to one of the two universal DNA segments;

    wherein;

    the 3′

    portion of the first universal primer anneals to the first universal segment;

    a 5′

    portion of the first universal primer comprises a nucleic acid sequence substantially identical to a portion of the first universal DNA segment, a portion of the second universal DNA segment, or a portion of the first target nucleic acid;

    an amplicon generated upon extension of the first universal primer forms an intramolecular hairpin stem between the 5′

    portion of the first universal primer and a portion of the amplicon complementary to the first universal DNA segment, the second universal DNA segment, or the first target nucleic acid;

    or wherein the first universal primer forms a circular structure upon annealing to the first tagged nucleic acid that brings the 3′ and

    5′

    -ends of the first universal primer close to each other; and

    ,formation of the hairpin stem or circular structure results or causes a change in a first detectable signal, the first detectable signal indicating the presence or quantity of the first target nucleic acid in the sample.

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