| 摘要 |
<p num="1"><br/><br/><br/>A method and assay kit for determination of thymidine kinase (TK) activity in <br/>a biological sample, such as blood, serum, plasma, Cerebral Spinal Fluid <br/>(CSF), pleural fluid, ascites, tissues, cells and extracts thereof, is <br/>described. The method comprises contacting, in a buffer, a Basic Reaction <br/>Mixture comprising: solid surface-attached primer and/or template, a modified <br/>deoxy nucleoside, such as BromodeoxyUridine, IododeoxyUridine, Fluorodeoxy- <br/>Uridine or VinyldexoyThymidine as a kinase enzyme substrate, a phosphate <br/>donor, a nucleotide polymerizing enzyme, and a kinase enzyme source devoid of <br/>TK activity, such as a yeast extract, with the biological sample. After <br/>incubation the amount of modified deoxy nucleoside that has been incorporated <br/>into the solid surface-attached primer and/or template, is determined and the <br/>TK activity present in the biological sample is directly proportional to the <br/>amount of incorporated modified deoxy nucleoside. The method and assay kit are <br/>useful in the diagnosing, prognosis monitoring of disease progression and <br/>treatment effects of cell-proliferation disorders or diseases, such as cancer, <br/>and in the screening of compounds, e.g. new drug candidates, affecting <br/>enzymatic pathways, which may obstruct the formation of thymidine phosphates <br/>or interfere with nucleic acid synthesis.<br/> |
