EP0519066A1 - Mesure de la production bacterienne de co 2? dans un fluorophore isole par controle d'un changement de fluorescence regule par absorption - Google Patents

Mesure de la production bacterienne de co 2? dans un fluorophore isole par controle d'un changement de fluorescence regule par absorption

Info

Publication number
EP0519066A1
EP0519066A1 EP92904836A EP92904836A EP0519066A1 EP 0519066 A1 EP0519066 A1 EP 0519066A1 EP 92904836 A EP92904836 A EP 92904836A EP 92904836 A EP92904836 A EP 92904836A EP 0519066 A1 EP0519066 A1 EP 0519066A1
Authority
EP
European Patent Office
Prior art keywords
fluorescence
absorbance
dye
sensor
blood culture
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP92904836A
Other languages
German (de)
English (en)
Other versions
EP0519066A4 (en
Inventor
Roger James Morris
Shoshana Bascomb
Jamie Bobolis
David Sherman
Carolyn Shelchuk Olson
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dade Behring Inc
Original Assignee
Baxter Diagnostics Inc
Dade MicroScan Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Baxter Diagnostics Inc, Dade MicroScan Inc filed Critical Baxter Diagnostics Inc
Publication of EP0519066A1 publication Critical patent/EP0519066A1/fr
Publication of EP0519066A4 publication Critical patent/EP0519066A4/en
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
    • G01N21/80—Indicating pH value
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
    • G01N21/272—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration for following a reaction, e.g. for determining photometrically a reaction rate (photometric cinetic analysis)

Definitions

  • This invention relates to a method to detect the presence or determine the concentration of microorganisms in a solution by regulating light reaching or exiting from a fluorophore encapsulated in or isolated by a chemically inert light transparent matrix.
  • Microorganisms present in bodily fluid can be detected using a culture bottle.
  • a culture bottle is a flask allowing positive cultures to be detected rapidly.
  • the flask is generally a transparent closed container filled with nutrient that promotes the growth of the organism.
  • bacteria in blood can be detected in culture.
  • microorganisms in a culture bottle have been detected by use of external detectors such as a magnifying lens, U.S. Patent No. 4,543,907 (Freudlich). Additionally, interal detectors such as liquid level indicators can show bacterial growth as a function of increased pressure in the vessel. Swaine et al., EPA 124,193 (Swaine et al.). Additionally, microorganisms can be detected by measuring changes in pH caused by bacterial growth, Mariel, G.B. Patent No. 1,601,689.
  • Still another method to detect microorganisms involves the use of a culture media that contains a compound which changes color or appearance according to the growth of microorganisms.
  • the change in the media can be detected with a spectrophotometer.
  • a spectrophotometer There are many examples of reactions used in Microbiology that rely on a color change. Bascomb, Enzyme Tests in Bacterial Identification. 19 Meth. Microbio. 105 (1987).
  • Bascomb Enzyme Tests in Bacterial Identification. 19 Meth. Microbio. 105 (1987).
  • Fermentation of carbohydrates results in the production of acid which causes a decrease in pH. This drop in pH can be easily detected by including a pH indicator like bromothymol blue or phenol red.
  • Chemical and enzymatic reactions are used to detect or quantitate the presence of certain substances in microbiological or other assays. Many of these tests rely on the development or change of color or fluorescence to indicate the presence or quantity of the substance of interest.
  • Another approach to determine if an organism can degrade a particular substrate is to use a reagent which is capable of reacting with one or more of the intermediates or final products. For example, the detection of the reduction of nitrate to nitrite. If nitrite is formed, then a pink to deep red color will result when sulfanilic acid and alpha-naphthylamine are added to the reaction mixture.
  • a synthetic ana]OR f a natural substrate to directly indicate the presence of an enzyme.
  • methylene blue can be reduced under certain conditions by the action of reductase, resulting in a shift from blue to colorless.
  • the oxidase assay relies on the interaction of cytochrome oxidase with N, N, N', N'-tetramethyl-p-phenylene- diamine producing a blue color.
  • Another example is the ability of microorganisms to degrade sulfur-containing amino acids as indicated by the production of S.
  • the organism is incubated with a high concentration of a sulfur-containing substrate (e.g. cysteine, cystine) in an acid environment.
  • a sulfur-containing substrate e.g. cysteine, cystine
  • the production of S is indicated by the formation of a black precipitate in the presence of ferric ammonium citrate.
  • Enzymes can usually act on more than one substrate. This allows for the use of synthetic enzyme substrates for the detection of enzyme activities.
  • Synthetic substrates contain a metabolic moiety conjugated with a chromatic or fluorescent moiety.
  • the conjugated molecule usually has a different absorption and/or emission spectrum from the unconjugated form.
  • the unconjugated chromatic or fluorescent moiety shows a considerably higher absorption or fluorescence coefficients than those of the conjugated molecule. This allows the measurement of small amounts of products of enzyme activities in the presence of the large amounts of conjugated substrate required for maximal enzyme activity.
  • An example of a synthetic enzyme substrate is o-nitro-phenol- ⁇ -galactopyranoside used for the detection of activity of the enzyme ⁇ -galactosidase.
  • the conjugated substrate is colorless.
  • the ⁇ -galactosidase enzyme hydrolyzes the substrate to yield ⁇ -galactosidase and o-nitro- phenol.
  • o-nitro-phenol absorbs strongly at 405nm, and its release can be measured by the increase in absorbance at that wavelength.
  • the monitoring of color or color end-product in chemical and microbial reactions is usually achieved in either of two ways; 1) the detection of color or color end-product can be achieved by visual observation and estimated qualitatively, or 2) the detection of color end-products or loss of color can be achieved by measuring the intensity of color instrumentally.
  • Spectrophotometers that measure light absorbance are commonly used for this purpose.
  • Shaffer U.S. Patent No. 4,495,293
  • Shaffer filed a patent application disclosing a method to fluorometrically determine a ligand in an assay solution using conventional fluorometric techniques.
  • the intensity of the fluorescence emitted by the assay solution is related to the change in transmissive properties of the assay solution produced by the interaction of the ligand to be determined and a reagent system capable of producing change in the transmissive properties of the assay solution in the presence of the ligand.
  • Shaffer discloses a method to monitor absorbance using a fluorophore in solution with the chromophore. In this method the fluorophore may interact with the assay cocktail and produce changes in fluorescence intensity which are unrelated to the change being measured.
  • the selection of the fluorophores is also restricted, in that pH dependent or environment sensitive fluorophores cannot be utilized.
  • Beggs & Sand, EPA 91,837 disclosed a solution based method for determination of tryptophan-deaminase activity by measuring the reduction in fluorescence in the presence of a chromophore produced by the interaction between indole pyruvic acid and metal ions using a fluorophore "whose fluorescence is capable of being quenched by the indole pyruvate-metal ion complex, the ions of the fluorophore being present throughout the incubation period".
  • Sands, U.S. Patent No.4,798,788 discloses a process to detect a nitrate reducing microorganism by measuring reduction of fluorescence in solution by causing the diazotization of the fluorophore.
  • a specific fluorophore needs to be chosen for each test to ensure that it will fluoresce under the conditions of the test, e.g. only few fluorophores fluoresce at pH of less than 2.0.
  • This invention provides a method to use a fluorophore enclosed or embedded in a chemically inert matrix which is transparent at the wavelengths of interest.
  • the fluorophore positioned to intersect the transmission light path, indirectly monitors absorbance or changes in the absorbance of a chromophore encapsulated or isolated by a gas permeable polymetric matrix.
  • the use of a fluorophore encapsulated in or isolated by a matrix allows for the sequential influence of reaction components on the intensity of light detected.
  • this invention relates to a multi-layer body fluid culture sensor comprised of a pH sensitive absorbance based dye spectrally coupled to a pH insensitive, or pH sensitive dye that is highly buffered, fluorescence based dye.
  • the pH sensitive absorbance based dye is encapsulated or isolated in a polymeric layer that is permeable to CO2 and water, but impermeable to protons.
  • the pH insensitive fluorophore is encapsulated or isolated in the second polymeric layer that may or may not be permeable to CC. and water.
  • This type of sensor may be used to detect or determine the concentration of microorganisms in bodily fluid.
  • this sensor is used to monitor microbial infections grown in a fluid culture bottle.
  • this sensor can be used to monitor bacterial growth. As bacteria grow they generate CO2. The ⁇ " ⁇ » ⁇ onerated by the bacteria diffuses into the polymeric layi-r - ⁇ r is in direct contact with a hydrated pH sensitive . ⁇ -' inc.- based dye.
  • the gas permeable, proton impermeable polymeric matrix is silicone. Additionally, in one embodiment of this invention a detector, such as a photomultiplier tube, is placed under the blood culture bottle to detect fluorescent emission.
  • FIG.l shows a schematic diagram of a multi-layer blood culture sensor.
  • FIG.2 shows a blood culture growth curve detected by a xylenol blue-rhodamine 101 sensor.
  • FIG.3 shows a blood culture growth curve detected by xylenol blue in silicone-rhodamine B in acrylic sensor.
  • FIG.4 shows a blood culture growth curve for a xylenol blue in silicone-6213 acrylic sensor.
  • FIG.5 shows a blood culture growth curve for- a bromothymol blue in silicone-rhodamine 101 in silicone sensor. Detailed Description - Best Mode
  • fluorescence from a fluorophore embedded in an inert light-transparent matrix is modulated by a pH sensitive absorbance dye embedded in a polymeric gas permeable, but proton impermeable matrix.
  • the assay is carried out in a blood culture bottle.
  • a fluorometric based colorimetric assay the fluorescence intensity is regulated by changes in absorbance of an interfering chromophore.
  • the chromophoric material alters the amount of light reaching the fluorophore and/or the amount of emitted light reaching the detector.
  • Spectrally compatible fluorescent and colorimetric indicators are selected so that as the pH changes due to the production of CO- by microorganisms present in the sample, the colorimetric indicators regulate the amount of light reaching the fluorophore and/or photodetector and, thus cause a change in the excitation and/or emission of the fluorescent dye. This change is detected with a fluorescent reader and can be correlated with the presence or concentration of microorganism in the sample.
  • a bodily fluid culture sensor is comprised of a pH sensitive absorbance based dye in or isolated by a polymeric gas permeable, but proton impermeable matrix, and a fluorescent dye in a second polymeric matrix.
  • Spectrally compatible fluorescent and colorimetric indicators are selected so that when an organism is present in a sample, the colorimetric indicator will regulate the amount of light reaching the fluorophore thereby causing a change in the emission intensity from the fluorescence dye reaching the photodetector.
  • the change indicating the presence of bacteria, is detected with a fluorometric reader.
  • spectrally compatible fluorescence and absorbance dyes are selected so that as carbonic acid is produced (CO2 and H2O) , the absorbance of the dye will change thereby regulating the amount of light reaching the fluorophore and/or photodetector, thus producing a change in the measured fluorescence. This change is detected with a fluorescence reader.
  • Spectrally compatible dyes are rhodamine b and xylenol blue. Additionally, proroothymol blue and rhodamine 101 are also spectrally compatible.
  • a culture bottle containing the appropriate growth media can be inoculated with E. col'i . As the organism grows, it produces CO2 gas. The silicone is permeable to the CO2.
  • the CO2 diffuses to the absorbance layer and reacts with water to produce carbonic acid ( CO ).
  • the carbonic acid causes a drop in the pH in the absorbance dye environment resulting in a change in measured absorbance. For example, as the pH drops in an absorbance layer containing the dye xylenol blue, the absorbance of xylenol blue decreases, thereby allowing more light to reach the fluorophore to excite it and thus increase the amount of fluorescence emitted at 590nm. A positive culture using xylenol blue as the absorbance dye is detected by increase the amount of fluorescence emitted at 590nm.
  • a positive culture using xylenol blue as the absorbance dye is detected by a measured increase in fluorescence as the xylenol blue decreases in absorbance.
  • the pH sensitive absorbance based dye is encapsulated in or isolated by a polymeric matrix that is gas permeable, but proton impermeable.
  • the polymeric matrix must be optically transparent in the visible region, permeable to gas, autoclavable, stable for at least six months, and proton impermeable.
  • silicone may function as the polymeric matrix used to encapsulate or isolate the absorbance based dye. Silicones found to meet these criteria were Dow, Rhone Poulenc, G.E. and Wacker.
  • the fluorescence based dyes can also be encapsulated in a polymeric matrix.
  • the polymeric matrix used for the fluorophore does not have to meet all of the above requirements listed for the matrix used to encapsulate or isolate the absorbance dye.
  • the similar features that it must possess are that it must be optically transparent in the visible region, autoclavable and stable for at least six months.
  • the polymeric matrix containing or isolating the absorbance based dye must be coupled to the polymeric matrix containing the fluorescent dye. It should be noted that the polymeric matrices must be in close proximity so that light that has been regulated by the absorbance layer will have an effect on the emission intensity of the fluorophore as received by the photodetector.
  • a bodily fluid culture sensor In the present invention, a bodily fluid culture sensor
  • FIG.l is comprised of a pH sensitive absorbance based dye encapsulated in or isolated by a polymeric gas permeable, but proton impermeable matrix 4 and a fluorescent dye in a second polymeric matrix 2.
  • Reflective surface 6 can be included to facilitate the transmission of light to the detecting element 12.
  • interrogation light enters the sensor and is regulated by pH sensitive matrix 2 which in turn causes a change in the fluorescence emission 10 of the fluorophore in matrix 4.
  • This sensor offers the advantage of maximal surface area.
  • an acrylic encapsulated fluorophore or silicone embedded fluorescence material is adhered to an absorbance dye isolating polymeric layer, to make a two layer sensor.
  • both the fluorescence and absorbance embedded material are poured into blood culture bottles.
  • the fluorophore embedded silicone material is poured on top of absorabnce embedded silicone.
  • the optical interrogation system comprises a visible output, 400-700nm, light source focused onto one end of a bifurcated fiber optic cable.
  • the common end is positioned close to the sensor, while the other end is positioned close to a photodetector, typically a photomultiplier tube.
  • Appropriate excitation and emission filters are used to select wavelengths of choice for each dye.
  • a beam splitter is used to divert a portion of the excitation light to a second photodetector and acts as a reference.
  • a photodetector converts light to a current source which is converted to a voltage using an operational amplifier.
  • a 12 bit analog to digital conversion offers sufficient dynamic range to read the voltage.
  • a computer program is then used to read, plot and store data.
  • a measurement is taken by first reading reference light intensity. Next the reading from the sensor disk is measured. The data is plotted by taking the ratio of reference, excitation light, to sample.
  • the absorbance of the absorbance dye changes, thereby changing the amount of light reaching the fluorescence layer and/or photodetector. This causes a change in emitted fluorescence that is detected.
  • the following examples serve to illustrate the method of the present invention. The concentration of reagents and other variable parameters are only shown to exemplify the methods of the present invention and are not to be considered limitation thereof.
  • Example 1 Xylenol Blue - Rhodamine 101 Sensor
  • Wacker silicone elastomer 3601 part A is thoroughly mixed with Wacker 3601 catalyst part B in a 9:1 ratio, as recommended by the manufacturer.
  • the absorbance layer mixture is then poured into an aluminum square mold to a thickness of 30/1000 of an inch and cured at 55 ° C for 2 hours.
  • Wacker silicone is prepared, as described above. Next 2% w/w of 7.5mM Rhodamine 101, in 50mK Tris-HCl buffer pH 8.5 in 95 ethylene glycol, is added to the silicone. The mixture is poured over the previously cured xylenol blue layer in the mold, described above, and cured at 55°C overnight. This cured, dehydrated, double layer sensor consists of two distinct layers, each 30/1000 of an inch thick. Disks may now be punched out of the mold and adhered onto the base of bottles using more silicone, ensuring that the absorbance layer is face down. Finally, the bottles are cured at 55 * C for 15 minutes, rehydrated with normal saline and autoclaved on the wet cycle for 17 minutes. Saline is replaced with growth media and inoculated with E. coli by injecting a suspension with a sterile needle through the septum. The blood culture bottle is placed in the instrument and fluorescence emission is measured.
  • Rhone Poulenc silicone elastomer 141 part A is thoroughly mixed with Rhone Poulenc 141 catalyst part B in a 10:1 ratio, as recommended by the manufacturer.
  • the absorbance layer mixture is then poured into an aluminum square mold to a thickness of 30/1000 of an inch. The mold is allowed to sit out on the countertop at room temperature for about one hour or until the bubbles have disappeared, at which time the mold is placed in the incubator to cure at 55°C for two hours.
  • Rhone-Poulenc silicone is prepared, as described above. Next, a 40/1,000" thick acrylic disc (Glasflex, Inc.). approximately 1 cm in diameter, containing 0.2 grams/lb of rhodamine B (Sigma) is glued onto the above absorbance layer using the Rhone-Poulenc silicone at the 10/1 ratio as glue. The double layer sensor is then placed back in the 55°C incubator for two hours to allow for adherence of the two layers. Following the curing, the double layer sensor is punched out with a cork borer, and glued onto the base of a Wheaton bottle, ensuring that the absorbance layer is face down, using the Rhone Poulenc silicone as mentioned above. The bottle is placed in the 55 " C incubator to cure for at least two hours. The bottle is then rehydrated overnite and tested the following day as described in Example 1.
  • Wacker silicone elastomer 3601 part A is thoroughly mixc. with Wacker 3601 catalyst part B in a 9:1 ratio, as recommended with Wacker 3601 catalyst part B in a 9:1 ratio, as reco ⁇ mended by the manufacturer.
  • Wacker silicone elastomer 3601 part A is thoroughly mixc. with Wacker 3601 catalyst part B in a 9:1 ratio, as recommended with Wacker 3601 catalyst part B in a 9:1 ratio, as reco ⁇ mended by the manufacturer.
  • 5% w/w of a 50mM xylenol blue, dissolved in 5mM borate buffer pH 11 containing 1% Tween 80, is added to the silicone and homogenized to ensure a uniform distribution of the dye.
  • the absorbance layer mixture is then poured into an aluminum square mold to a thickness of 30/1000 of an inch and cured at 55 * C for two hours.
  • a 40/1,000" thick acrylic disc (Glasflex, Inc.), approximately 1 cm in diameter, referred to as No. 6213 Red (Glassflex Standard Product) is glued onto the above absorbance layer using the Wacker silicone at the 9/1 ratio as glue.
  • the double layer sensor is then placed back in the 55 * C incubator for two hours to allow for adherence of the two layers.
  • the double layer sensor is punched out with a cork borer, and glued onto the base of a Wheaton bottle, ensuring that the absorbance layer is face down, using the Rhone Poulenc silicone as mentioned above.
  • the bottle is placed in the 55 * C incubator to cure for at least two hours.
  • the bottle is then rehydrated overnite and tested the following day as described in Example 1.
  • Wacker silicone elastomer 3601 part A is thoroughly mixed with Wacker 3601 catalyst part B in a 9:1 ratio, as recommended by the manufacturer.
  • the absorbance layer mixture is then poured into an aluminum square mold to a thickness of 30/1000 of an inch and cured at 55°C for two hours.
  • Wacker silicone is prepared, as described above. Next 2% w/w of 7.5mM Rhodamine 101, in 50mM Tris-HCl buffer pH 8.5 in 95% ethylene glycol, is added to the silicone. The mixture is poured over the previously cured xylenol blue layer in the mold, described above to isolate the absorbance layer. This sensor is then cured at 55 " C overnight. This cured, dehydrated, double layer sensor consists of two distinct layers, each 30/1000 of an inch thick. Disks may now be punched out of the mold and adhered onto the base of bottles using more silicone, ensuring that the absorbance layer is face down.

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  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Biotechnology (AREA)
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  • Molecular Biology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Theoretical Computer Science (AREA)
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  • Genetics & Genomics (AREA)
  • Plasma & Fusion (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
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Abstract

Capteur multicouche pour cultures de sang comprenant un colorant à absorption sensible au pH encapsulé dans ou isolé par une première matrice transparente perméable au gaz, imperméable aux protons, ainsi qu'un colorant à fluorescence insensible au pH dans une seconde matrice (4), les première et seconde matrices étant couplées de manière spectrale. On décrit également un procédé d'utilisation de ce capteur afin de détecter des micro-organismes dans un flacon de mise en culture de sang.
EP19920904836 1991-01-04 1991-12-23 Measurement of bacterial co 2? production in an isolated fluorophore by monitoring an absorbance regulated change of fluorescence Ceased EP0519066A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US63848191A 1991-01-04 1991-01-04
US638481 1991-01-04

Publications (2)

Publication Number Publication Date
EP0519066A1 true EP0519066A1 (fr) 1992-12-23
EP0519066A4 EP0519066A4 (en) 1993-08-18

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EP19920904836 Ceased EP0519066A4 (en) 1991-01-04 1991-12-23 Measurement of bacterial co 2? production in an isolated fluorophore by monitoring an absorbance regulated change of fluorescence

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EP (1) EP0519066A4 (fr)
JP (1) JPH05504263A (fr)
AU (1) AU652423B2 (fr)
CA (1) CA2077560A1 (fr)
NO (1) NO923436L (fr)
WO (1) WO1992012413A1 (fr)

Families Citing this family (7)

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JPH07252367A (ja) * 1994-03-16 1995-10-03 Dow Corning Kk オルガノポリシロキサン樹脂をマトリックス材料とする機能性素子の製造方法
US5611900A (en) * 1995-07-20 1997-03-18 Michigan State University Microbiosensor used in-situ
AT405103B (de) * 1996-10-16 1999-05-25 Avl Verbrennungskraft Messtech Sensorschicht zur quantitativen bestimmung zumindest einer chemischen komponente einer gasförmigen oder flüssigen probe
GB0419325D0 (en) * 2004-09-01 2004-09-29 Perkinelmer Ltd A method of analysing a sample including fluorescent labels and apparatus therefor
US8512975B2 (en) * 2008-07-24 2013-08-20 Biomerieux, Inc. Method for detection and characterization of a microorganism in a sample using time dependent spectroscopic measurements
ITNA20090042A1 (it) * 2009-07-02 2011-01-03 Maurizio Baldassarre Un nuovo metodo ottico per una determinazione rapida e semplice di analiti
US11460410B2 (en) * 2019-04-08 2022-10-04 Puma SE Bioindicator component applied to an article

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0214768A2 (fr) * 1985-08-08 1987-03-18 The Regents Of The University Of California Optrode d'absorption et d'émission
EP0283116A2 (fr) * 1987-02-06 1988-09-21 Shiley Incorporated Système de palpeur
US4822746A (en) * 1986-06-25 1989-04-18 Trustees Of Tufts College Radiative and non-radiative energy transfer and absorbance modulated fluorescence detection methods and sensors
EP0352610A2 (fr) * 1988-07-25 1990-01-31 Abbott Laboratories Capteurs physiologiques à fibre optique

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MX153322A (es) * 1979-05-23 1986-09-12 Miles Lab Dispositivo analitico quimioluminiscente mejorado para detectar un componente en una muestra de liquidos corporales
US4803049A (en) * 1984-12-12 1989-02-07 The Regents Of The University Of California pH-sensitive optrode
US4867919A (en) * 1986-10-10 1989-09-19 Minnesota Mining And Manufacturing Company Method of making a gas sensor
US4851195A (en) * 1987-08-17 1989-07-25 Pfizer Hospital Products Group, Inc. Carbon dioxide sensor
US4945060A (en) * 1988-03-15 1990-07-31 Akzo N. V. Device for detecting microorganisms
US5173434A (en) * 1990-11-05 1992-12-22 Baxter Diagnostics Inc. Measurement of color reactions by monitoring a change of fluorescence

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0214768A2 (fr) * 1985-08-08 1987-03-18 The Regents Of The University Of California Optrode d'absorption et d'émission
US4822746A (en) * 1986-06-25 1989-04-18 Trustees Of Tufts College Radiative and non-radiative energy transfer and absorbance modulated fluorescence detection methods and sensors
EP0283116A2 (fr) * 1987-02-06 1988-09-21 Shiley Incorporated Système de palpeur
EP0352610A2 (fr) * 1988-07-25 1990-01-31 Abbott Laboratories Capteurs physiologiques à fibre optique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO9212413A1 *

Also Published As

Publication number Publication date
EP0519066A4 (en) 1993-08-18
JPH05504263A (ja) 1993-07-08
CA2077560A1 (fr) 1992-07-05
NO923436D0 (no) 1992-09-03
AU1263892A (en) 1992-08-17
WO1992012413A1 (fr) 1992-07-23
NO923436L (no) 1992-09-03
AU652423B2 (en) 1994-08-25

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