EP0441469A1
Expired / Lapsed
EP
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Immunospecific and bioluminescent assay of cellular ATP
Assignee: Promega Corp
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Filing: 1991-01-02
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Granted: —
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Expires: 2011-01-02
Abstract
Abstract The detection of living organisms through the use of a bioluminescent assay for a living cell detection metabolite such as ATP with an antibody-like mediated cell capture onto a solid phase is described. The assay utilizes detection methods after immunospecific cell capture to a solid phase. The method involves cell capture, removal of non-specific or other contaminating materials, cell lysis and detection of the detection metabolite utilizing a light producing enzyme such as the firefly-luciferase enzyme technique. The invention is also directed to kits for performing such detections.
Inventors
- 👤 Terry M. Sivesind
- 👤 Randall L. Dimond
- 👤 James W. Schumm
- 👤 Edward E. Pahuski
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 05:42
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
115 words
A process for determining the presence of an antigenic cell substance including a detection metabolite in a sample, comprising: a) contacting the sample with cell binding molecules designed to bind to the antigenic cell substance, the cell binding molecules being bound to a solid carrier insoluble in the sample, to form a complex between the cell binding molecules and antigenic cell substance; b) separating the solid carrier from the sample; c) lysing the cell wall of the antigenic cell substance to release the detection metabolite; d) adding a light producing enzyme reagent to the treated sample of step c); and e) detecting the presence of the detection metabolite in the treated sample of step d).
Claim 2
Dependent Claim
12 words
A process according to claim 1 wherein the detection metabolite is ATP.
Claim 3
Dependent Claim
16 words
A process according to claim 1 wherein the detection metabolie is NAD⁺, NADH, FMNH₂ or FMN.
Claim 4
Dependent Claim
28 words
A process according to any one of claims 1 to 3 wherein the light-producing enzyme is a firefly-luciferase reagent or click beetle luciferase reagent or a variation thereof.
Claim 5
Dependent Claim
16 words
A process according to claim 4 wherein the light producing enzyme is a bacterial firefly-luciferase reagent.
Claim 6
Dependent Claim
31 words
A process according to any one of claims 1 to 5 wherein the sample is a food, liquid, body fluid or a solution derived from a food, liquid or body fluid.
Claim 7
Dependent Claim
25 words
A process according to any one of the preceding claims wherein the antigenic cell substance is a prokaryotic cellular organism or an eukaryotic cellular organism.
Claim 8
Dependent Claim
17 words
A process according to any one of the preceding claims wherein the cell binding molecules are antibodies.
Claim 9
Dependent Claim
15 words
A process according to claim 8 wherein the antibodies are polyclonal antibodies or monoclonal antibodies.
Claim 10
Dependent Claim
34 words
A process according to any one of the preceding claims wherein the solid carrier is a microtiter dish, test tube, polystyrene beads, plastic beads, a resin, paper, a plastics material or a magnetic particle.
Claim 11
Dependent Claim
23 words
A process according to any one of claims 1 to 10 wherein the solid carrier is an ELISA plate or ELISA test tube.
Claim 12
Dependent Claim
36 words
A process according to claim 10 wherein the solid carrier is a magnetic particle coated with avidin or a magnetic particle coated with streptavidin and the antibodies are cell specific antibodies labeled with biotin or iminobiotin.
Claim 13
Dependent Claim
18 words
A process according to claim 10 wherein the solid carrier is a magnetic particle coated with protein A.
Claim 14
Dependent Claim
22 words
A process according to any one of claims 1 to 9 wherein the solid carrier is a dipstick coated with the antibodies.
Claim 15
Dependent Claim
23 words
A process according to any one of claims 1 to 14 werehin the cell binding proteins are directly attched to the solid carrier.
Claim 16
Dependent Claim
42 words
A process according to any one of claims 1 to 14 wherein the cell binding proteins are attached to the solid carrier by chemical means or by bridging molecules which are biotin, avidin, streptavidin, secondary antibodies, lectins, protein A or protein G.
Claim 17
Dependent Claim
29 words
A process according to any one of the preceding claims wherein the unreacted antigen is separated from the solid carrier by washing the solid carrier with a buffer solution.
Claim 18
Dependent Claim
14 words
A process according to claim 17 wherein the buffer solution is phosphate buffered saline.
Claim 19
Dependent Claim
22 words
A process according to any one of the preceding claims wherein the cell walls are lysed by the addition of an acid.
Claim 20
Dependent Claim
19 words
A process according to claim 19 wherein the acid is nitric acid, trichloroacetic acid, perchloric acid or sulphuric acid.
Claim 21
Dependent Claim
23 words
A process according to any of claims 1 to 18 wherein the cell walls are lysed by the addition of an organic solvent.
Claim 22
Dependent Claim
25 words
A process according to any one of claims 1 to 18 wherein the cell walls are lysed by the addition of a detergent or surfactant.
Claim 23
Dependent Claim
59 words
A process according to any one of the preceding claims wherein the concentration of the antigenic cell substance is the sample is determined wherein the emitted light is measured in step e) and which comprises: f) calculating the concentration of the detection metabolite from the measurement or emitted light; and g) calculating the antigenic cell concentration in the sample.
Claim 24
Dependent Claim
17 words
A process according to claim 23 for determining the presence and concentration of Salmonella in a sample.
Claim 25
Dependent Claim
71 words
A diagnostic kit for the detection of the presence of an antigenic cell substance in a sample, comprising: a) a container containing a cell binding molecule against the antigenic cell substance coupled to a water insoluble solid carrier; b) a container a buffer solution; c) a container containing a cell lysing or detection metabolite releasing solution; and d) a container containing assay reagents to perform an assay for the detection metabolite.
Claim 26
Dependent Claim
27 words
A kit according to claim 25 which has one or more of the features of claims 2 to 5, 7 to 16, 18 to 20 and 24.
References Cited (Backward Citations)
US3660240A
Cited
US3745090A
Cited
US4385113A
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EP0016552A1
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US4282287A
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DE3224484A1
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EP0153283A1
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WO1986007094A1
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WO1989002929A1
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EP0363510A1
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