EP0238352A2
Expired / Lapsed
EP
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Method for the selective detection of microbial nucleotides
Assignee: 3M Co
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Filing: 1987-03-20
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Granted: —
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Expires: 2007-03-20
Abstract
Abstract A method for the selective determination of microbial cells, in a sample suspected of containing both microbial and non-microbial cells, is described which involves the selective release and enzymatic inactivation of non-microbial nucleotides, followed by the rapid and specific inhibition of the inactivating enzyme and the release and detection of microbial nucleotides in an appropriate assay, such as a bioluminescent assay.
Inventors
- 👤 John G.H.M. Vossen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 15:11
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/48; C12Q1/02; C12Q1/04; C12Q1/66
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
109 words
A method for the selective determination of microbial nucleotides that is useful for a sample suspected of containing both non-microbial and microbial cells, characterized in that said method comprises the steps of: (1) selectively releasing non-microbial nucleotides, (2) inactivating substantially all of the released non-microbial nucleotides by use of an effective amount of an inactivating enzyme, other than a hydrolase, which can be inhibited by a specific inhibitor, (3) inhibiting substantially all of the inactivating enzyme by use of an effective amount of a specific inhibitor, (4) selectively releasing microbial nucleotides, and (5) detecting a statistically significant amount of the released microbial nucleotides by means of an appropriate assay.
Claim 2
Independent Claim
109 words
A method for the selective determination of microbial nucleotides that is useful for a sample suspected of containing both non-microbial and microbial cells, characterized in that said method comprises the steps of: (1) selectively releasing non-microbial nucleotides, (2) inactivating substantially all of the released non-microbial nucleotides by use of an effective amount of an inactivating enzyme, other than a hydrolase, which can be inhibited by a specific inhibitor, (3) inhibiting substantially all of the inactivating enzyme by use of an effective amount of a specific inhibitor, (4) selectively releasing microbial nucleotides, and (5) detecting a statistically significant amount of the released microbial nucleotides by means of an appropriate assay.
Claim 3
Dependent Claim
28 words
A method according to claim 1 further characterized in that the time required to inactivate the released non-microbial nucleotides is on the order of ten minutes or less.
Claim 4
Dependent Claim
52 words
A method according to claim 1 further characterized in that the inhibitor is selected from the group consisting of N-ethylmaleimide, organomercurials, iodoacetate, iodoacetamide, and aryl halides; the inactivating enzyme is a phosphotransferase selected from the group consisting of glycerokinase, hexokinase and adenylate kinase and the microbial nucleotides detected are i adenosine triphosphate.
Claim 5
Dependent Claim
16 words
A method according to claim 1 further characterized in that the assay is a bioluminescent assay.
Claim 6
Dependent Claim
31 words
A method according to claim 1 further characterized in that the non-microbial nucleotides are released by a non-ionic releasing agent, and the microbial nucleotides are released by an ionic releasing agent.
Claim 7
Dependent Claim
21 words
A method according to claim 1 further characterized in that the release and inactivation of non-microbial nucleotides are carried out simultaneously.
Claim 8
Dependent Claim
26 words
A method according to claim 1 further characterized in that the inhibition of the inactivating enzyme and the release of microbial nucleotides are carried out simultaneously.
Claim 9
Dependent Claim
22 words
A method according to claim 1 further characterized in that the inactivating enzyme is glycerokinase and the specific inhibitor is p-chloromercuriphenylsulfonic acid.
Claim 10
Dependent Claim
29 words
A kit for carrying out the method of claim 1 characterized in that said kit comprises predetermined and individually packaged amounts of inactivating enzyme and its corresponding specific inhibitor.
References Cited (Backward Citations)
No citation records indexed.
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