Method of amplifying ATP and use thereof
|Posted date||Sep 6, 2011|
|Country||United States of America|
|Date of filing||Apr 28, 2010|
|Gazette Date||Dec 9, 2010|
|Gazette Date||Aug 23, 2011|
|International application number||JP2004011186|
|International publication number||WO2005010208|
|Date of international filing||Jul 27, 2004|
|Date of international publication||Feb 3, 2005|
|Title||Method of amplifying ATP and use thereof|
The ATP amplification method 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.
Also provided is 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||
1. A kit for detecting the presence of a microorganism, comprising an adenosine triphosphate amplification reagent containing 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; and an adenosine triphosphate detection reagent for detecting 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 and wherein the fusion protein is not present in a complex with adenosine diphosphate.
2. The kit of claim 1, further comprising a cell lysis reagent.
3. A fusion protein that has a polyphosphate kinase and an adenylate kinase in this order from the N-terminal and that has been subjected to an apyrase treatment and a pyrophosphate treatment so as to remove adenosine diphosphate bound to the fusion protein, wherein the fusion protein is not present in a complex with adenosine diphosphate.
|IPC(International Patent Classification)||
|Reference ( R and D project )||PRESTO Conversion and Control by Advanced Chemistry AREA|
Contact Information for " Method of amplifying ATP and use thereof "
- Japan Science and Technology Agency Department of Intellectual Property Management
- URL: http://www.jst.go.jp/chizai/
- Address: 5-3, Yonbancho, Chiyoda-ku, Tokyo, Japan , 102-8666
- Fax: 81-3-5214-8417