Patent ID: 7141377

Claim:
A kit for detecting a nucleic acid sequence in a sample comprising: (a) a set of oligonucleotide primers, wherein a first primer contains a sequence complementary to a region in one strand of the nucleic acid sequence and primes the synthesis of a first extension product, and a second primer contains a sequence complementary to a region in said first extension product and primes the synthesis of a nucleic acid strand complementary to said first extension product, and wherein each oligonucleotide primer is selected to anneal to its complementary template upstream of any labeled oligonucleotide annealed to the same nucleic acid strand, (b) at least one labeled oligonucleotide containing a sequence complementary to a region of the nucleic acid sequence, wherein said labeled oligonucleotide anneals within the nucleic acid sequence bounded by the oligonucleotide primers of part (a) and wherein said labeled oligonucleotide is blocked at the 3′ terminus to prohibit incorporation of said labeled oligonucleotide into a primer extension product, wherein said blocking is achieved by adding a chemical moiety to the 3′ hydroxyl of the last nucleotide, which moiety does not also serve as a label for subsequent detection or by removing said 3′-hydroxyl, and wherein said labeled oligonucleotide is provided in a soluble form and it generates a signal concurrently with the amplification of said nucleic acid sequence using said set of oligonucleotide primers.