Patent ID: 7282355
Filing Date: 2007-10-16
Classification: C12Q

Abstract:
1. A method for detecting the presence of a target nucleic acid molecule in a plurality of templates, comprising: (a) combining (i) a detection probe comprising a universal sequence; (ii) a first oligonucleotide comprising a 3′ terminal sequence complementary to a first portion of a known nucleotide sequence of a target nucleic acid molecule and a second sequence complementary to the universal sequence; (iii) a second oligonucleotide comprising a 5′ terminal sequence complementary to a second portion of the known nucleotide sequence of the target nucleic acid molecule, wherein the second portion of the known nucleotide sequence of the target nucleic acid molecule is adjacent to the first portion of the known nucleotide sequence; (iv) a primer comprising a 3′ terminal sequence complementary to a portion of the second oligonucleotide, wherein the 3′ terminal sequence of the primer is not complementary to the second portion of the known nucleotide sequence of the target nucleic acid molecule; and (v) a plurality of templates suspected of containing the target nucleic acid molecule, wherein the combining is under conditions wherein complementary sequences hybridize to one another; (b) incubating the product of step (a) with a ligase under conditions wherein the ligase forms a covalent bond between the 3′ terminus of the first oligonucleotide hybridized to the first portion of the known nucleotide sequence and the 5′ terminus of the second oligonucleotide hybridized to the second portion of the known nucleotide sequence; (c) incubating the product of step (b) with a DNA polymerase under conditions wherein the primer is extended; and (d) detecting the presence of a non-hybridized detection probe, wherein the non-hybridized detection probe is dissociated from the product of step (b) as a result of an activity of the DNA polymerase, wherein the presence of the non-hybridized detection probe indicates the presence of the target nucleic acid molecule in the plurality of templates.