Patent ID: 7129045

Claim:
A method for detecting an analyte comprising: a) performing a specific binding pair reaction for detecting the analyte wherein polynucleotide kinase is present as a label on a specific binding pair member wherein the specific binding pair reaction comprises binding polynucleotide kinase labeled specific binding partner with its cognate specific binding partner for purposes of detecting the analyte, wherein the labeled specific binding partner is the analyte, an analog of the analyte, or a substance having a specific binding affinity for the analyte; b) reacting polynucleotide kinase, a nucleotide triphosphate and an oligonucleotide having a 5′-OH group and comprising a sequence S2 to phosphorylate oligonucleotide S2 at the 5′-OH group; c) providing a reaction mixture comprising i) oligonucleotide S2, ii) a single stranded nucleic acid template comprising contiguous sequence regions C1 and C2, iii) a first oligonucleotide having a sequence S1 which is hybridized to region C1 of the template, and v) a ligase and a cofactor for the ligase; d) ligating with the ligase oligonucleotide S1 and oligonucleotide 5′-phosphate S2 to form a ligation product, wherein the ligation only occurs if oligonucleotide S2 is phosphorylated by polynucleotide kinase; e) detecting the ligated oligonucleotide product of step d as indicating the presence or activity of the polynucleotide kinase; and f) relating the activity of the polynucleotide kinase to the amount of the analyte.