US3933592A
Expired / Lapsed
US
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Method of detecting living microorganisms
Assignee: Labcorp Early Development Laboratories Inc
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Filing: 1970-10-05
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Granted: 1976-01-20
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Expires: 1990-10-05
Abstract
Abstract The presence or absence of living microorganisms is determined by treating a sample containing said microorganisms with an adenosine triphosphate reactive mixture containing luciferin, luciferase and a cation in the presence of oxygen and measuring the light emission produced.
Inventors
- 👤 John R. Clendenning
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 01:02
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/66; C12Q1/02; C12Q1/18; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/83; Y10S435/831; Y10S435/832; Y10S435/834; Y10S435/859; Y10S435/88; Y10S435/881; Y10S435/882; Y10S435/884; Y10S435/911; Y10S435/94; Y10S435/942
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
97 words
A method for testing an unknown sample of material, said sample containing substantially no adenosine triphosphate which is not in living microorganisms therein, and said material being suspected to contain living microorganisms which comprises: a. treating said sample to render adenosine triphosphate in any living microorganisms present therein available for reaction, b. contacting said sample with an adenosine triphosphate-reactive mixture comprising luciferin, luciferase, and a cation in the presence of oxygen, and c. measuring the light emission from any ensuing reaction, said light emission being indicative of the presence and amount of living microorganisms in said sample.
Claim 2
Independent Claim
97 words
A method for testing an unknown sample of material, said sample containing substantially no adenosine triphosphate which is not in living microorganisms therein, and said material being suspected to contain living microorganisms which comprises: a. treating said sample to render adenosine triphosphate in any living microorganisms present therein available for reaction, b. contacting said sample with an adenosine triphosphate-reactive mixture comprising luciferin, luciferase, and a cation in the presence of oxygen, and c. measuring the light emission from any ensuing reaction, said light emission being indicative of the presence and amount of living microorganisms in said sample.
Claim 3
Dependent Claim
10 words
A method of claim 1 wherein said microorganisms are bacteria.
Claim 4
Dependent Claim
30 words
A method of claim 1 wherein the sample comprises a solid material and the treatment of said sample includes the step of forming an aqueous suspension of said solid material.
Claim 5
Dependent Claim
39 words
A method of claim 1 wherein the treatment of said sample includes the step of reacting said sample with a phosphorylating enzyme to convert any adenosine triphosphate precursors which are present to adenosine triphosphate prior to measuring light emission.
Claim 6
Dependent Claim
25 words
A method of claim 1 wherein the treatment of said sample includes the step of extracting adenosine triphosphate present in said microorganisms from said microorganisms.
Claim 7
Dependent Claim
11 words
A method of claim 5 wherein the extraction is with dimethylsulfoxide.
Claim 8
Dependent Claim
12 words
A method of claim 5 wherein the extraction is with perchloric acid.
Claim 9
Dependent Claim
12 words
A method of claim 5 including the step of ultrasonic cell disruption.
Claim 10
Dependent Claim
10 words
A method of claim 1 wherein said cation is magnesium.
Claim 11
Dependent Claim
23 words
A method of claim 1 wherein adenosine triphosphate which is not in living microorganisms is removed from said sample prior to step (a).
Claim 12
Dependent Claim
10 words
A method of claim 10 wherein removal is by washing.
Claim 13
Dependent Claim
10 words
A method of claim 10 wherein removal is by centrifuging.
Claim 14
Independent Claim
185 words
A method for testing an unknown sample of material, said sample containing substantially no adenosine triphosphate which is not in living microorganisms therein, and said material being suspected to contain living microorganisms which comprises: a. treating said sample to render adenosine triphosphate in any living microorganisms present therein available for reaction, b. contacting said sample with an adenosine triphosphate-reactive mixture comprising luciferin, luciferase, and a cation in the presence of oxygen, and c. measuring the light emission from any ensuing reaction, said light emission being indicative of the presence and amount of living microorganisms in said sample, said method including the step of incubating a second such approximately equal sample of said material in a nutrient medium, repeating the process of steps (a), (b), and (c) on said incubated sample, and comparing the light emission produced by the unincubated sample with that produced by the incubated sample, the presence of living microorganisms in said material being indicated by emission of light in the initial test and confirmed by an increase in light emission produced by said incubated sample over that produced by said unincubated sample.
Claim 15
Independent Claim
185 words
A method for testing an unknown sample of material, said sample containing substantially no adenosine triphosphate which is not in living microorganisms therein, and said material being suspected to contain living microorganisms which comprises: a. treating said sample to render adenosine triphosphate in any living microorganisms present therein available for reaction, b. contacting said sample with an adenosine triphosphate-reactive mixture comprising luciferin, luciferase, and a cation in the presence of oxygen, and c. measuring the light emission from any ensuing reaction, said light emission being indicative of the presence and amount of living microorganisms in said sample, said method including the step of incubating a second such approximately equal sample of said material in a nutrient medium, repeating the process of steps (a), (b), and (c) on said incubated sample, and comparing the light emission produced by the unincubated sample with that produced by the incubated sample, the presence of living microorganisms in said material being indicated by emission of light in the initial test and confirmed by an increase in light emission produced by said incubated sample over that produced by said unincubated sample.
Claim 16
Dependent Claim
76 words
A method of claim 13 for detecting the presence of a specific type of living microorganism suspected to be present in an unknown sample of material wherein said incubation is in a nutrient medium and under conditions favorable to the growth of said suspected type of microorganism, the presence of said suspected type of living microorganism being indicated by an increase in light emission produced by said incubated sample over that produced by said unincubated sample.
Claim 17
Dependent Claim
20 words
A method of claim 13 wherein adenosine triphosphate which is not in living microorganisms is removed prior to step (a).
Claim 18
Dependent Claim
9 words
A method claim 15 wherein removal is by washing.
Claim 19
Dependent Claim
145 words
A method of claim 1 for identifying the type of antibiotic effective for treatment of living microorganisms suspected to be present in an unknown sample of material which includes the steps of providing a plurality of equal aliquots of a nutrient broth for living microorganisms suspected to exist in said sample, adding a different antibiotic to separate aliquots, introducing equal portions of said sample into said aliquots, and incubating said separate aliquots, test steps (a), (b), and (c) being applied to aliquots initially and after incubation, the presence of living microorganisms in said material being indicated by emission of light by an aliquot in said initial test, and the effectiveness of a particular antibiotic being indicated by failure of an incubated aliquot to exhibit a significant increase in light emission after incubation as compared with an aliquot which produced light emission in the initial test.
Claim 20
Dependent Claim
159 words
A method of claim 1 for identifying the type of living microorganism suspected to be present in an unknown sample of material which includes the steps of providing a plurality of equal aliquots of a nutrient broth for living microorganisms suspected to exist in said sample, adding a different antibody to each of separate aliquots, said antibodies being specific for different microorganisms, introducing equal portions of said sample into said aliquots, and incubating said separate aliquots, test steps (a), (b), and (c) being applied to aliquots initially and after incubation, the presence of living microorganisms in said material being indicated by emission of light by an aliquot in said initial test and failure of a particular incubated aliquot to exhibit a significant increase in light emission after incubation, as compared with an aliquot which produced light emission in the initial test, being indicative that the antibody contained in that incubated aliquot is specific for microorganisms contained in said material.
References Cited (Backward Citations)
US2905592A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-01-19 | 1979-07-21 | 1980-01-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-01-19 | 1983-07-21 | 1984-01-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-01-19 | 1987-07-21 | 1988-01-20 | $7700.00 | Window Closed / Lapsed |