Patent ID: 7521188

Claim:
A method of detecting a target DNA in a sample, the method comprising: performing a DNA amplification process on the sample, wherein the DNA amplification process uses an amplification mixture including at least an intact labeled oligonucleotide, a DNA polymerase enzyme, and the target DNA, wherein the target DNA is from phakopsora pachyrhizi; exposing the intact labeled oligonucleotide in the amplification mixture to exonuclease activity of the DNA polymerase enzyme in the amplification mixture, the intact labeled oligonucleotide including a site-specific sequence and a co-factor conjugated to the site-specific sequence, thereby hydrolyzing the intact labeled oligonucleotide and releasing a fragment of the oligonucleotide including the co-factor, wherein the site specific sequence is 5′-amino-hexyl-CCAAAAGGTACACCTGTTTGAGTCA-3′ (SEQ ID NO: 1), and the co-factor is pyrroloquinoline quinone; combining the fragment with an apo-enzyme, wherein combining the fragment including the co-factor with the apo-enzyme produces an enzymatic signal that is optically detectable; and optically detecting the enzymatic signal, thereby detecting the target DNA.