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Method of amplifying ATP and use thereof meetings achieved

Foreign code F110005470
File No. K05323WO
Posted date Sep 6, 2011
Country United States of America
Application number 56695704
Gazette No. 20080213754
Gazette No. 7745160
Date of filing Jul 27, 2004
Gazette Date Sep 4, 2008
Gazette Date Jun 29, 2010
International application number JP2004011186
International publication number WO2005010208
Date of international filing Jul 27, 2004
Date of international publication Feb 3, 2005
Priority data
  • P2003-202992 (Jul 29, 2003) JP
  • 2004WO-JP11186 (Jul 27, 2004) WO
Title Method of amplifying ATP and use thereof meetings achieved
Abstract (US7745160)
The ATP amplification method of the present invention is a method for amplifying and detecting a very trace amount of exogenous ATP by allowing a fusion protein (PPK-ADK) of a polyphosphate kinase and an adenylate kinase, the fusion protein not containing ADP, to act on a mixture of ATP, AMP, and a polyphosphate compound.
The present invention also provides an ultrasensitive ATP amplification method by which ATP at a single cell level can be amplified and detected, and an ultrasensitive microbial assay based on this ATP amplification method.
Scope of claims [claim1]
1. A method for amplifying adenosine triphosphate comprising: allowing a fusion protein that has a polyphosphate kinase and an adenylate kinase in this order from the N-terminal to act on a mixture containing adenosine triphosphate, adenosine monophosphate, and a polyphosphate compound thereby amplifying adenosine triphosphate in the mixture, wherein the fusion protein has been subjected to an apyrase treatment and a pyrophosphate treatment so as to remove adenosine diphosphate bound to the fusion protein.
[claim2]
2. A method for detecting adenosine triphosphate comprising: allowing a fusion protein that has a polyphosphate kinase and an adenylate kinase in this order from the N-terminal to act on a mixture of adenosine triphosphate, adenosine monophosphate, and a polyphosphate compound thereby amplifying adenosine triphosphate in the mixture; and
detecting the amplified adenosine triphosphate,
wherein the fusion protein has been subjected to an apyrase treatment and a pyrophosphate treatment so as to remove adenosine diphosphate bound to the fusion protein.
[claim3]
3. A method for detecting the presence of a microorganism comprising: treating a sample containing a microorganism to prepare a sample containing adenosine triphosphate;
adding the sample containing adenosine triphosphate to an adenosine triphosphate amplification system thereby amplifying adenosine triphosphate in the sample;
detecting the amplified adenosine triphosphate; and
detecting the presence of the microorganism by the presence of the detected adenosine triphosphate,
wherein the adenosine triphosphate amplification system comprises adenosine monophosphate, a polyphosphate compound, and a fusion protein that has a polyphosphate kinase and an adenylate kinase in this order from the N-terminal,
wherein the fusion protein has been subjected to an apyrase treatment and a pyrophosphate treatment so as to remove adenosine diphosphate bound to the fusion protein.
  • Inventor, and Inventor/Applicant
  • KURODA AKIO
  • CALPIS
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) PRESTO Conversion and Control by Advanced Chemistry AREA
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