Patent ID: 8216810

Claim:
A method for determining the presence or absence of at least one target nucleic acid sequence in a test sample, the method comprising: (a) contacting a detecting system, which comprises at least one detecting subsystem, wherein the at least one detecting subsystem comprises: at least one capture nucleic acid comprising 40 to 70 consecutive variably degenerate nucleotides complementary to the target nucleic acid sequence or at least one capture nucleic acid comprising consecutive nucleotides complementary to the target nucleic acid sequence, wherein the capture nucleic acid binds to a first region in the target nucleic acid sequence, and at least one detector agent which comprises: (i) a binder nucleic acid comprising 40 to 70 consecutive variably degenerate nucleotides complementary to the target nucleic acid sequence, wherein the binder nucleic acid binds to a second region in the target nucleic acid sequence, and (ii) a reporter nucleic acid comprising 50 to 350 nucleotides which is linked to the binder nucleic acid, wherein the reporter nucleic acid further comprises a first primer binding sequence and a second primer binding sequence, wherein the first and the second primer binding sequences are different; with the test sample, under conditions suitable for formation of a complex comprising the target nucleic acid sequence, the capture nucleic acid, and the detector agent and forming a mixture; (b) immobilizing the capture nucleic acid in the mixture on a solid support; (c) washing the solid support to remove unbound detector agent and unbound target nucleic acid and isolate the complex comprising the target nucleic acid sequence, the capture nucleic acid, and the detector agent if the target nucleic acid is present in the test sample; (d) adding a first primer which binds to the first primer binding sequence of the reporter nucleic acid and a second primer which binds to a sequence complementary to the second primer binding sequence of the reporter nucleic acid onto the solid support in step (c) and amplifying the reporter nucleic acid or a portion thereof, using the first primer and the second primer to produce an amplified product, wherein detecting the amplified product indicates the presence of the target nucleic acid sequence in the test sample.