- 专利标题: Terminal-phosphate-labeled nucleotides and methods of use
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申请号: US10113030申请日: 2002-04-01
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公开(公告)号: US07052839B2公开(公告)日: 2006-05-30
- 发明人: John Nelson , Carl Fuller , Anup Sood , Shiv Kumar
- 申请人: John Nelson , Carl Fuller , Anup Sood , Shiv Kumar
- 申请人地址: US NJ Piscataway
- 专利权人: Amersham Biosciences Corp
- 当前专利权人: Amersham Biosciences Corp
- 当前专利权人地址: US NJ Piscataway
- 代理商 Royal N. Ronning, Jr.; Yonggang Ji
- 主分类号: C12Q1/68
- IPC分类号: C12Q1/68
摘要:
The present invention describes methods of detecting a nucleic acid in a sample, based on the use of terminal-phosphate-labeled nucleotides as substrates 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.