Patent ID: 8632974
Filing Date: 2014-01-21
Classification: C12Q

Abstract:
1. A method for assessing a sample for target nucleic acid, said method comprising: (a) contacting said sample with a probe nucleic acid comprising an amplifying restriction endonuclease and a nucleotide sequence complementary to a sequence of said target nucleic acid under conditions wherein, if said target nucleic acid is present in said sample, at least a portion of said target nucleic acid hybridizes to at least a portion of said probe nucleic acid to form a double-stranded portion of nucleic acid comprising a restriction endonuclease cut site, (b) contacting said double-stranded portion of nucleic acid with a recognition restriction endonuclease having the ability to cut said double-stranded portion of nucleic acid at said restriction endonuclease cut site under conditions wherein said recognition restriction endonuclease cleaves said double-stranded portion of nucleic acid at said restriction endonuclease cut site, thereby separating a portion of said probe nucleic acid comprising an amplifying restriction endonuclease from at least another portion of said probe nucleic acid, (c) contacting said portion of said probe nucleic acid comprising an amplifying restriction endonuclease with a first nucleic acid comprising an amplifying restriction endonuclease and a double-stranded portion of nucleic acid comprising a restriction endonuclease cut site of the amplifying restriction endonuclease of said portion of said probe nucleic acid comprising an amplifying restriction endonuclease under conditions wherein the amplifying restriction endonuclease of said portion of said probe nucleic acid comprising an amplifying restriction endonuclease cleaves said first nucleic acid at said restriction endonuclease cut site of the amplifying restriction endonuclease of said portion of said probe nucleic acid comprising an amplifying restriction endonuclease, thereby separating a portion of said first nucleic acid comprising an amplifying restriction endonuclease from at least another portion of said first nucleic acid, (d) contacting said portion of said first nucleic acid comprising an amplifying restriction endonuclease with a reporter nucleic acid comprising a double-stranded portion of nucleic acid comprising a restriction endonuclease cut site of the amplifying restriction endonuclease of said portion of said first nucleic acid comprising an amplifying restriction endonuclease under conditions wherein the amplifying restriction endonuclease of said portion of said first nucleic acid comprising an amplifying restriction endonuclease cleaves said reporter nucleic acid at said restriction endonuclease cut site of the amplifying restriction endonuclease of said portion of said first nucleic acid comprising an amplifying restriction endonuclease, thereby separating a portion of said reporter nucleic acid from at least another portion of said reporter nucleic acid, and (e) determining the presence or absence of said portion of said reporter nucleic acid, wherein the presence of said portion of said reporter nucleic acid indicates that said sample contains said target nucleic acid, and wherein the absence of said portion of said reporter nucleic acid indicates that said sample does not contain said target nucleic acid.