Patent ID: 8048633

Claim:
A method of performing a nucleic acid amplification assay comprising: (a) combining reagents, DNA polymerase, a target nucleic acid, and a modified primer, said modified primer comprising a sequence of bases complementary to a first region of said target nucleic acid, a polymerase blocking region, a single stranded hybridization region attached to said polymerase blocking region, and a detectable label, said primer predominantly having a secondary structure in solution in a lower temperature range and not in an elevated temperature range, wherein said secondary structure involves annealing one or more self-complementary regions between portions of said primer region and portions of said single stranded hybridization region, wherein said modified primer is capable of priming said target nucleic acid and hybridizing with an oligonucleotide complementary to said single stranded hybridization region at a first elevated temperature range, and substantially not in a second lower temperature range; (b) cycling the mixture of (a) at said first temperature range to provide multiple copies of an amplicon incorporating said modified primer, wherein said incorporated primer is capable of hybridizing with an oligonucleotide complementary to said single stranded hybridization region in said first elevated temperature range and in said second lower temperature range; (c) reducing the temperature of the mixture to the second temperature range; (d) exposing said mixture to a capture oligonucleotide complementary to said single stranded hybridization region; (e) hybridizing said single stranded hybridization region of said amplicon incorporating said modified primer, with said capture oligonucleotide; and (f) detecting said label associated with said hybridization.