发明名称 |
TERMINAL-PHOSPHATE-LABELED NUCLEOTIDES AND METHODS OF USE |
摘要 |
The present invention relates to improved methods of detecting a target using a labeled substrate or substrate analog. The methods comprise reacting the substrate or substrate analog in an enzyme-catalyzed reaction which produces a labeled moiety with independently detectable signal only when such substrate or substrate analog reacts. The present invention, in particular, describes methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrate for nucleic acid polymerases. The methods provided by this invention utilize a nucleoside polyphosphate, dideoxynucleoside polyphosphate, or deoxynucleoside polyphosphate analogue which has a colorimetric dye, chemiluminescent, or fluorescent moiety, a mass tag or an electrochemical tag attached to the terminal-phosphate. When a nucleic acid polymerase uses this analogue as a substrate, an enzyme-activatable label would be present on the inorganic polyphosphate by-product of phosphoryl transfer. Cleavage of the polyphosphate product of phosphoryl transfer via phosphatase leads to a detectable change in the label attached thereon. When the polymerase assay is performed in the presence of a phosphatase, there is provided a convenient method for real-time monitoring of DNA or RNA synthesis and detection of a target nucleic acid. |
申请公布号 |
WO2004072297(A3) |
申请公布日期 |
2007.01.18 |
申请号 |
WO2004US02785 |
申请日期 |
2004.01.30 |
申请人 |
AMERSHAM BIOSCIENCES CORP;FULLER, CARL;KUMAR, SHIV;SOOD, ANUP;NELSON, JOHN |
发明人 |
FULLER, CARL;KUMAR, SHIV;SOOD, ANUP;NELSON, JOHN |
分类号 |
C12Q1/68;C07H19/10;C07H19/20;C07H21/00;C12Q;C12Q1/00;C12Q1/42;G01N33/52;G01N33/58 |
主分类号 |
C12Q1/68 |
代理机构 |
|
代理人 |
|
主权项 |
|
地址 |
|