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METHODS AND COMPOSITIONS FOR DETECTING TARGET NUCLEIC ACIDS

  • US 20130005594A1
  • Filed: 05/17/2012
  • Published: 01/03/2013
  • Est. Priority Date: 03/29/2010
  • Status: Active Grant
First Claim
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1. A method for detecting a plurality of different target nucleic acids in a sample, wherein each target nucleic acid comprises a first target domain adjacent to a second target domain and a third target capture domain located upstream or downstream from said first and second target domains, said method comprising:

  • (a) providing a plurality of ligation substrates each comprising;

    (i) one of said target nucleic acids;

    (ii) a first set of ligation probes comprising;

    (A) a first nucleic acid ligation probe comprising;

    (1) a first probe domain hybridized to a first target domain of said one target nucleic acid sequence;

    (2) a 5′

    -ligation moiety; and

    (B) a second nucleic acid ligation probe comprising;

    (1) a second probe domain hybridized to a second target domain of said one target nucleic acid sequence; and

    (2) a 3′

    ligation moiety;

    (b) ligating said first and second ligation probes without the use of a ligase enzyme to form a first plurality of ligation products;

    (c) hybridizing target capture probes comprising a capture moiety to said third target domain of said target nucleic acids to form target complexes;

    (d) capturing said target complexes on a surface using said capture moiety;

    (e) amplifying said ligation products to form amplicons;

    (f) detecting said amplicons,thereby detecting said target nucleic acids.

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