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EP0126019A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

High resolution method of measuring ATP, and concentrating and measuring unicellular organisms

Assignee: Revvity Health Sciences Inc
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Filing: 1984-05-15
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Granted: —
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Expires: 2004-05-15
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Abstract

Abstract A method of concentrating and measuring the amount of cells in a sample is described. A-sample containing the plurality of cells is passed upwardly or laterally through a series of membrane filters and concentrated on the filter. The filter membrane containing the concentrated cell is then sequentially treated with lysing agent to release the adenosine triphosphate from the organism and a luminescent reagent to react with the released adenosine triphosphate. A luminescent response is thereby produced directly on the filter membrane and this luminescent response is measured with a luminometer. Not only does this method provide an accurate way of detecting very minute quantities of cells but it provides a relatively quick and easy method of concentrating and measuring cells in large volumes of biological or industrial samples which contain low levels of organisms. This method can also be used to identify and measure sources of adenosine triphosphate. Another aspect of the disclosed method is the utilization of a series of filters, each of which traps and concentrates cells of different size. For example, a sample containing a plurality of cell types can be passed through a series of filter membranes, with different pore size each of which allows passage of all but one unicellular organism, resulting in a concentration of different cells on different filter membranes. The filter membranes can then be processed and measured on the luminometer as described above.

Inventors

  • 👤 Leroy J. Everett
  • 👤 Tenlin Sie Tsai
  • 👤 Norma J. Ramey

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-10-31 15:33
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04

Claims Analysis

Total Claims: 10
Claim 1
Independent Claim 78 words
A method of separating microbial ATP, non-microbial ATP, and extra-cellular ATP, comprising passing a sample containing said ATP upwards through a series of filter membranes of decreasing pore size, all membranes permeable to the extra-cellular ATP, at least one filter membrane impermeable to cells containing the microbial ATP, and at least one filter membrane impermeable to cells containing non-microbial ATP, the filter membrane impermeable to non-microbial ATP being upstream of the filter membrane impermeable to the microbial ATP.
Claim 2
Independent Claim 78 words
A method of separating microbial ATP, non-microbial ATP, and extra-cellular ATP, comprising passing a sample containing said ATP upwards through a series of filter membranes of decreasing pore size, all membranes permeable to the extra-cellular ATP, at least one filter membrane impermeable to cells containing the microbial ATP, and at least one filter membrane impermeable to cells containing non-microbial ATP, the filter membrane impermeable to non-microbial ATP being upstream of the filter membrane impermeable to the microbial ATP.
Claim 3
Independent Claim 54 words
A method of separating a plurality of cell types contained in a single sample comprising passing the sample upwards through a series of filter membranes of decreasing pore size, each filter having a different pore size from the other filters and each filter sequentially being permeable to all but one of the unicellular organisms.
Claim 4
Independent Claim 54 words
A method of separating a plurality of cell types contained in a single sample comprising passing the sample upwards through a series of filter membranes of decreasing pore size, each filter having a different pore size from the other filters and each filter sequentially being permeable to all but one of the unicellular organisms.
Claim 5
Independent Claim 96 words
A method of seperating and measuring cellular ATP and extra-cellular ATP comprising passing a sample containing said ATP uowards through a series of filter membranes of decreasing pore size, all membranes permeable to the extra-cellular ATP, at least one filter membrane impermeable to cells containing the cellular ATP, treating the impermeable filter membrane containing the cells sequentially with lysing agent to release the adenosime triphosphate from the cells and a luminescent reagent to react with the released ATP thereby producing a luminescent response directly on the filter membrane and measuring the luminescent response with a luminometer.
Claim 6
Independent Claim 96 words
A method of seperating and measuring cellular ATP and extra-cellular ATP comprising passing a sample containing said ATP uowards through a series of filter membranes of decreasing pore size, all membranes permeable to the extra-cellular ATP, at least one filter membrane impermeable to cells containing the cellular ATP, treating the impermeable filter membrane containing the cells sequentially with lysing agent to release the adenosime triphosphate from the cells and a luminescent reagent to react with the released ATP thereby producing a luminescent response directly on the filter membrane and measuring the luminescent response with a luminometer.
Claim 7
Independent Claim 50 words
A method of isolating substantially pure bacteria cells from a sample also containing non-microbial soluble interferences comprising passing a sample containing said cells upwards through a series of filter membranes of decreasing pore size, all membranes permeable to the interferences and at least one filter membrane impermeable to the cells.
Claim 8
Independent Claim 50 words
A method of isolating substantially pure bacteria cells from a sample also containing non-microbial soluble interferences comprising passing a sample containing said cells upwards through a series of filter membranes of decreasing pore size, all membranes permeable to the interferences and at least one filter membrane impermeable to the cells.
Claim 9
Independent Claim 59 words
A method of cell enrichment of a specific cell type comprising concentrating the specific cell type from a sample containing a plurality of cell types by passing said cell sample containing the plurality of cell types upwards through a series of filter membranes of decreasing pore size, at least one filter membrane being impermeable to the specific cell type.
Claim 10
Independent Claim 59 words
A method of cell enrichment of a specific cell type comprising concentrating the specific cell type from a sample containing a plurality of cell types by passing said cell sample containing the plurality of cell types upwards through a series of filter membranes of decreasing pore size, at least one filter membrane being impermeable to the specific cell type.

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