US5567598A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Microbial monitoring device
Assignee: Becton Dickinson and Co
•
Filing: 1993-03-03
•
Granted: 1996-10-22
•
Expires: 2013-03-03
Abstract
Abstract This invention presents methods for detection and evaluation of metabolic activity of microorganisms based upon their ability to consume dissolved oxygen. The methods utilize a fluorescence detection system which makes use of the sensitivity of the fluorescent emission of certain compounds to the presence of oxygen, which quenches (diminishes) the compound's fluorescent emission in a concentration dependent manner. Respiring microorganisms will affect the oxygen concentration of a liquid medium in which they are immersed. Thus, this invention provides a convenient system to gather information on the presence, identification and metabolic activity of microorganisms by determining their effect on the oxygen concentration of the media in which they are present.
Inventors
- 👤 David T. Stitt
- 👤 Gregory J. Burrell
- 👤 Kwok-Yu Hu
- 👤 James F. Monthony
- 👤 Robert Sapitowicz
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 01:42
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/18; C12Q2304/00; C12Q2304/40; C12Q2304/44; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S436/00; Y10S436/80
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
188 words
A method for detecting the presence of respiring microorganisms in a fluid comprising: (i) contacting said fluid with a sensor composition which comprises a fluorescent compound that exhibits a reduction in fluorescent intensity, when irradiated with light containing wavelengths which cause said compound to fluoresce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the microorganism; (ii) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iii) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (iv) comparing said measurement to that of a control not containing a respiring microorganism, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring microorganisms and a calculated threshold, wherein a change in fluorescent intensity relative to the fluorescent intensity of the control is indicative of the presence of respiring microorganisms; and (v) in the event that no such increase is measured or observed, repeat steps (ii), (iii), and (iv) as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 2
Independent Claim
188 words
A method for detecting the presence of respiring microorganisms in a fluid comprising: (i) contacting said fluid with a sensor composition which comprises a fluorescent compound that exhibits a reduction in fluorescent intensity, when irradiated with light containing wavelengths which cause said compound to fluoresce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the microorganism; (ii) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iii) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (iv) comparing said measurement to that of a control not containing a respiring microorganism, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring microorganisms and a calculated threshold, wherein a change in fluorescent intensity relative to the fluorescent intensity of the control is indicative of the presence of respiring microorganisms; and (v) in the event that no such increase is measured or observed, repeat steps (ii), (iii), and (iv) as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 3
Dependent Claim
14 words
The method of claim 1 where said matrix is a rubber or plastic matrix.
Claim 4
Dependent Claim
13 words
The method of claim 1 wherein said matrix is a silicone rubber matrix.
Claim 5
Dependent Claim
15 words
The method of claim 1 wherein said fluorescent compound is adsorbed on solid silica particles.
Claim 6
Dependent Claim
15 words
The method of claim 1 wherein said fluorescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) salt.
Claim 7
Dependent Claim
14 words
The method of claim 5 wherein said fluorescent compound is tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.
Claim 8
Dependent Claim
16 words
The method of claim 1 wherein said fluorescent compound is a tris-2, 2'-bipyridyl ruthenium (II) salt.
Claim 9
Dependent Claim
15 words
The method of claim 7 wherein said fluorescent compound is tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate.
Claim 10
Dependent Claim
13 words
The method of claim 1 wherein said fluorescent compound is 9, 10-diphenyl anthracene.
Claim 11
Dependent Claim
22 words
The method of claim 1 wherein said fluid is isolated from atmospheric oxygen wherein said fluid is contained in a closed system.
Claim 12
Dependent Claim
13 words
The method of claim 1 wherein said fluid is exposed to atmospheric oxygen.
Claim 13
Independent Claim
244 words
A method for determining the effects of an antibiotic or antimicrobial composition on a respiring microorganism comprising: (i) preparing a broth of said microorganism: (ii) contacting said broth with a sensor composition which comprises a fluorescent compound that exhibits a reduction in fluorescent intensity, when irradiated with light containing wavelengths which cause said compound to fluoresce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the microorganism; (iii) admixing with said broth, a quantity of said antibiotic or antimicrobial composition; (iv) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (v) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; and (vi) comparing said measurement to that of a control wherein said control is selected from the group consisting of: a reagent control not in contact with a respiring microorganism or the antibiotic or antimicrobial composition, a reagent control in contact with a respiring microorganism but not in contact with the antibiotic or antimicrobial composition and a calculated threshold, wherein a change in fluorescent intensity relative to the control is indicative of the susceptibility or resistivity of the microorganisms to the quantity of said antibiotic or antimicrobial composition; and (vii) in the event that no such change is measured or observed, repeat steps (iv), (v) and (vi), as needed, to determine the effects of the antibiotic or antimicrobial on the respiring microorganisms.
Claim 14
Independent Claim
244 words
A method for determining the effects of an antibiotic or antimicrobial composition on a respiring microorganism comprising: (i) preparing a broth of said microorganism: (ii) contacting said broth with a sensor composition which comprises a fluorescent compound that exhibits a reduction in fluorescent intensity, when irradiated with light containing wavelengths which cause said compound to fluoresce, upon exposure to oxygen, wherein the presence of the sensor composition is non-destructive to the microorganism; (iii) admixing with said broth, a quantity of said antibiotic or antimicrobial composition; (iv) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (v) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; and (vi) comparing said measurement to that of a control wherein said control is selected from the group consisting of: a reagent control not in contact with a respiring microorganism or the antibiotic or antimicrobial composition, a reagent control in contact with a respiring microorganism but not in contact with the antibiotic or antimicrobial composition and a calculated threshold, wherein a change in fluorescent intensity relative to the control is indicative of the susceptibility or resistivity of the microorganisms to the quantity of said antibiotic or antimicrobial composition; and (vii) in the event that no such change is measured or observed, repeat steps (iv), (v) and (vi), as needed, to determine the effects of the antibiotic or antimicrobial on the respiring microorganisms.
Claim 15
Dependent Claim
14 words
The method of claim 12 where said matrix is a rubber or plastic matrix.
Claim 16
Dependent Claim
16 words
The method of claim 12 wherein said rubber or plastic matrix is a silicone rubber matrix.
Claim 17
Dependent Claim
15 words
The method of claim 12 wherein said fluorescent compound is adsorbed on solid silica particles.
Claim 18
Dependent Claim
15 words
The method of claim 12 wherein said fluorescent compound is a tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) salt.
Claim 19
Dependent Claim
14 words
The method of claim 16 wherein said fluorescent compound is tris-4,7-diphenyl-1,10-phenanthroline ruthenium (II) chloride.
Claim 20
Dependent Claim
16 words
The method of claim 12 wherein said fluorescent compound is a tris-2, 2'-bipyridyl ruthenium (II) salt.
Claim 21
Dependent Claim
15 words
The method of claim 18 wherein said fluorescent compound is tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate.
Claim 22
Dependent Claim
13 words
The method of claim 12 wherein said fluorescent compound is 9, 10-diphenyl anthracene.
Claim 23
Dependent Claim
13 words
The method of claim 12 wherein said broth is isolated from atmospheric oxygen.
Claim 24
Dependent Claim
13 words
The method of claim 12 wherein said broth is exposed to atmospheric oxygen.
Claim 25
Dependent Claim
177 words
(amended) A method for detecting the presence of respiring microorganisms in a fluid comprising: (i) contacting said fluid with a fluorescent sensor composition which comprises a fluorescent compound selected from the group consisting of tris-4,7-diphenyl-1,10-phenanthroline-ruthenium (II) chloride and tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate, wherein the presence of the sensor composition is non-destructive to the microorganism; (ii) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iii) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (iv) comparing said measurement to that of a control, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring microorganisms and a calculated threshold, wherein a change in fluorescent intensity relative to the fluorescent intensity of the control is indicative of the presence of respiring microorganisms; and (v) in the event that no such increase is measured or observed, repeat steps (ii), (iii), and (iv), as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 26
Dependent Claim
177 words
(amended) A method for detecting the presence of respiring microorganisms in a fluid comprising: (i) contacting said fluid with a fluorescent sensor composition which comprises a fluorescent compound selected from the group consisting of tris-4,7-diphenyl-1,10-phenanthroline-ruthenium (II) chloride and tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate, wherein the presence of the sensor composition is non-destructive to the microorganism; (ii) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iii) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (iv) comparing said measurement to that of a control, wherein said control is selected from the group consisting of: a reagent control not in contact with respiring microorganisms and a calculated threshold, wherein a change in fluorescent intensity relative to the fluorescent intensity of the control is indicative of the presence of respiring microorganisms; and (v) in the event that no such increase is measured or observed, repeat steps (ii), (iii), and (iv), as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 27
Dependent Claim
14 words
The method of claim 23 where said matrix is a rubber or plastic matrix.
Claim 28
Dependent Claim
13 words
The method of claim 23 wherein said matrix is a silicone rubber matrix.
Claim 29
Dependent Claim
15 words
The method of claim 23 wherein said fluorescent compound is adsorbed on solid silica particles.
Claim 30
Dependent Claim
14 words
The method of claim 23 wherein said fluid compound is isolated from atmospheric oxygen.
Claim 31
Dependent Claim
14 words
The method of claim 23 wherein said fluid compound is exposed to atmospheric oxygen.
Claim 32
Independent Claim
231 words
A method of determining the effect of an antibiotic or antimicrobial composition on a respiring microorganism comprising: (i) preparing a broth of said microorganism: (ii) contacting said broth with a sensor composition which comprises a fluorescent compound selected from the group consisting of tris-4,7-diphenyl-1,10-phenanthroline-ruthenium (II) chloride and tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate, wherein the presence of the sensor composition is non-destructive to the microorganism; (iii) admixing with said broth a quantity of said antibiotic or antimicrobial composition: (iv) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iv) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (v) comparing said measurement to that of a control, wherein said control is selected from the group consisting of: a reagent control not in contact with a respiring microorganism or the antibiotic or antimicrobial composition, a reagent control in contact with a respiring microorganism but not in contact with the antibiotic or antimicrobial composition and a calculated threshold, wherein a change in fluorescent intensity relative to the control is indicative of the susceptibility or resistivity of the microorganisms to quantity of the antibiotic or antimicrobial composition: and (vi) in the event that no such change is measured or observed, repeat steps (iii), (iv), and (v), as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 33
Independent Claim
231 words
A method of determining the effect of an antibiotic or antimicrobial composition on a respiring microorganism comprising: (i) preparing a broth of said microorganism: (ii) contacting said broth with a sensor composition which comprises a fluorescent compound selected from the group consisting of tris-4,7-diphenyl-1,10-phenanthroline-ruthenium (II) chloride and tris-2,2'-bipyridyl ruthenium (II) chloride hexahydrate, wherein the presence of the sensor composition is non-destructive to the microorganism; (iii) admixing with said broth a quantity of said antibiotic or antimicrobial composition: (iv) irradiating said sensor composition with light containing wavelengths which cause said fluorescent compound to fluoresce; (iv) measuring or visually observing the fluorescent light intensity from said fluorescent compound while irradiating said sensor compound with said light; (v) comparing said measurement to that of a control, wherein said control is selected from the group consisting of: a reagent control not in contact with a respiring microorganism or the antibiotic or antimicrobial composition, a reagent control in contact with a respiring microorganism but not in contact with the antibiotic or antimicrobial composition and a calculated threshold, wherein a change in fluorescent intensity relative to the control is indicative of the susceptibility or resistivity of the microorganisms to quantity of the antibiotic or antimicrobial composition: and (vi) in the event that no such change is measured or observed, repeat steps (iii), (iv), and (v), as needed, to detect the presence of respiring microorganisms in said fluid.
Claim 34
Dependent Claim
14 words
The method of claim 29 where said matrix is a rubber or plastic matrix.
Claim 35
Dependent Claim
13 words
The method of claim 29 wherein said matrix is a silicone rubber matrix.
Claim 36
Dependent Claim
15 words
The method of claim 29 wherein said fluorescent compound is adsorbed on solid silica particles.
Claim 37
Dependent Claim
23 words
The method of claim 29 wherein said broth compound is isolated from atmospheric oxygen wherein said fluid is contained in a closed system.
Claim 38
Dependent Claim
14 words
The method of claim 23 wherein said fluid compound is exposed to atmospheric oxygen.
References Cited (Backward Citations)
US3509026A
Cited
US3612866A
Cited
US4643968A
Cited
GB2132348A
Cited
DE3346810A1
Cited
US5030420A
Cited
US4798788A
Cited
US4810655A
Cited
US4824789A
Cited
US4824789B1
Cited
EP0333253A2
Cited
EP0448923A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-10-22 | 2000-04-22 | 2000-10-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-10-22 | 2004-04-22 | 2004-10-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-10-22 | 2008-04-22 | 2008-10-22 | $7700.00 | Window Closed / Lapsed |