×

Methods and composition for detecting targets

  • US 20040235005A1
  • Filed: 10/23/2003
  • Published: 11/25/2004
  • Est. Priority Date: 10/23/2002
  • Status: Abandoned Application
First Claim
Patent Images

1. A method for detecting the presence or absence of at least one target nucleic acid sequence in a sample comprising:

  • forming a ligation reaction composition comprising the sample, and a ligation probe set for each target nucleic acid sequence, the probe set comprising (a) at least one first probe, comprising a target-specific portion and a 5′

    primer-specific portion, wherein the 5′

    primer-specific portion comprises a sequence, and (b) at least one second probe, comprising a target-specific portion and a 3′

    primer-specific portion, wherein the 3′

    primer-specific portion comprises a sequence, wherein the probes in each set are suitable for ligation together when hybridized adjacent to one another on a complementary target sequence;

    forming a test composition by subjecting the ligation reaction composition to at least one cycle of ligation, wherein adjacently hybridizing complementary probes are ligated to one another to form a ligation product comprising the 5′

    primer-specific portion, the target-specific portions, and the 3′

    primer-specific portion;

    forming at least one amplification reaction composition comprising;

    at least a portion of the test composition;

    a polymerase;

    a double-stranded-dependent specific label, wherein the double-stranded-dependent label has a first detectable signal value when the double-stranded-dependent label is not exposed to double-stranded nucleic acid; and

    at least one primer set, the primer set comprising (i) at least one first primer comprising the sequence of the 5′

    primer-specific portion of the ligation product, and (ii) at least one second primer comprising a sequence complementary to the sequence of the 3′

    primer-specific portion of the ligation product;

    subjecting the at least one amplification reaction composition to at least one amplification reaction; and

    detecting a second detectable signal value at least one of during and after the at least one amplification reaction, wherein a threshold difference between the first detectable signal value and the second detectable signal value indicates the presence of the target nucleic acid sequence, and wherein no threshold difference between the first detectable signal value and the second detectable signal value indicates the absence of the target nucleic acid sequence.

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×