WO1985000890A1 - Systeme chimique bioluminescent et methode de detection de la presence d'agents chimiques dans un milieu - Google Patents
Systeme chimique bioluminescent et methode de detection de la presence d'agents chimiques dans un milieu Download PDFInfo
- Publication number
- WO1985000890A1 WO1985000890A1 PCT/US1984/001217 US8401217W WO8500890A1 WO 1985000890 A1 WO1985000890 A1 WO 1985000890A1 US 8401217 W US8401217 W US 8401217W WO 8500890 A1 WO8500890 A1 WO 8500890A1
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- WIPO (PCT)
- Prior art keywords
- substrain
- sensitive
- resistant
- substrains
- chemical
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Classifications
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- 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
- C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- 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
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2520/00—Use of whole organisms as detectors of pollution
Definitions
- This invention relates to the detection of chemical agents, especially toxic substances, in a vapor, aerosol, or liquid medium or on solid surfaces. More particularly, this invention • concerns the use of genetically selected sensitive and resistant biolumi- nescent microorganism substrains to indicate the pres ⁇ ence of specific chemical agents by comparing the lumi ⁇ nescence of the sensitive and resistant microorganism substrains.
- the present invention obviates these inherent problems contained in prior detection methods by pro ⁇ viding a bioluminescent chemical system capable of detecting a plurality of chemical agents in either a vapor, aerosol, or liquid medium.
- the bioluminescent chemical system of the present invention is capable of detecting relatively minuscule levels of toxic agents even when relatively high concentrations of similar chemical agents or other chemical agents are present in the medium. Since the sensitive and resistant mutant substrains used in the present invention are geneti ⁇ cally derived from the same parental strain of biolumi ⁇ nescent microorganisms, the presence of other chemical agents does not affect the differences in light output of the selective and resistant substrains when exposed to the chemical agents the detection of which is sought.
- the bioluminescent chemical system of the present invention is capable of detecting the presence of one or more chemical agents in a vapor, aerosol, or liquid medium or on solid surfaces.
- the system includes one or more pairs of strains of photolumi- nescent microorganisms. Each pair of photoluminescent
- OMPI microorganisms comprises a genetically-selected sensi ⁇ tive substrain and a genetically-selected resistant substrain.
- the sensitive and resistant substrains are derived by mutation from a single parent strain.
- a comparator means such as a self-normalizing difference circuit or an electro-optic sensor, observes the dif ⁇ ferences in photoluminescence between the sensitive substrain and resistant substrain of each parent strain.
- An indicator means such.as an alarm-display circuit, registers the progressive change in the rela ⁇ tive light output of the sensitive and resistant sub ⁇ strains derived from the parent strain as observed by the comparator means.
- the sensitive sub ⁇ strain and resistant substrain of each parent are lyo- philized after mutation with the lyophilized substrains being activated just prior to use in detecting the presence of the chemical agents.
- a plurality of mutant pairs, each having a sensitive substrain sensitive to a specific chemical agent and a resistant substrain resistant to a specific chemical agent, may be used in the bioluminescent chemical system to detect the pres ⁇ ence of a plurality .of chemical agents contained in the medium.
- a storage and activation system for the bioluminescent detection of the presence of one or more chemical agents in a vapor, aerosol, or liquid medium is pro ⁇ vided and includes a sheet unit with one or more cap ⁇ sules formed within the sheet unit. Each capsule is divided into a microorganism compartment and an activa ⁇ tion solution compartment. Lyophilized microorganisms, which are capable of being photoluminescent when acti ⁇ vated, are encapsulated within each microorganism com ⁇ partment and an activation solution is encapsulated within each activation solution compartment.
- Crushing means such as a pair of rollers, are provided for breaking the capsules to allow the activation solution to come into contact with the lyophilized microorgan ⁇ isms contained within the microorganism compartment.
- the activation solution contacts the lyophilized microorganisms, the microorganisms are activated into a state of photoluminescence.
- the present invention provides a method of detecting one or more chemical agents in a vapor, aerosol, or liquid medium by the use of a photo ⁇ luminescent chemical system which includes the steps of deriving by mutation a genetically selected sensitive substrain and a genetically resistant substrain from a parent strain of a photoluminescent microorganism.
- the sensitive and resistant substrains are separately grown and then lyophilized.
- the lyophilized substrains are stored dry in capsules which protect the substrains from oxygen, water, and light.
- the lyophilized substrains are activated by adding an activation solution to the lyo ⁇ philized substrains.
- the sensitive substrain and resistant substrain are exposed to the vapor, aerosol, or liquid medium or solid surface containing the chemi ⁇ cal agents so that the luminescence of the sensitive substrains can be compared to the luminescence of the resistant substrains to detect and identify specific chemical agents.
- Figure 1 is a block diagram representing the present invention.
- Figure 2 is a perspective view of a preferred- embodiment of the storage and activation system of the present invention.
- O PI Figure 2a is a sectional view taken along 2-2 of Figure 2.
- Figure 3 is a side schematic view of the cap ⁇ sules containing lyophilized microorganisms used in the storage and activation system.
- Figure 4 is a side view of an alternative embodiment of the capsules used in the storage and activation system.
- Figure 5 is a perspective view of an alternative embodiment of the storage and activation system.
- Figure 6 is the schematic diagram of a simplified circuit system of the present invention.
- FIG. 1 is a block diagram of a biolumines ⁇ cent chemical system 10 in accordance with the present invention.
- the bioluminescent chemical system 10 includes a transportable unit 11 containing a sensitive substrain 12 and a resistant substrain 13.
- the sensi ⁇ tive substrain 12 and resistant substrain 13 are genet ⁇ ically selected and derived by mutation from the same parent strain of photoluminescent microorganisms.
- the sensitive substrain 12 is sensitive to the specific chemical agent contained in the aerosol, vapor, or liquid medium. This sensitivity is exhibited by the decreased luminescence of the sensitive substrain upon exposure to the specific chemical agent.
- the resistant substrain 13 remains relatively unaf- fected by exposure to the specific chemical agent to which the sensitive substrain is sensitive and, accord ⁇ ingly, the photoluminescence of the resistant substrain 13 does not significantly decrease upon exposure to that specific chemical agent.
- the detec- tion unit 11 has a power pack 14 which periodically
- O PI advances a fresh unit of sensitive substrain 12 and resistant- substrain 13 in the detection unit 11 to an operation position for exposure to the chemical agents in the medium.
- Electro-optic sensors 15 observe the differ ⁇ ences in light output between the sensitive substrain 12 and the resistant substrain 13, when both are exposed to the same chemical agents contained within the same medium. The differences in light output between the sensitive and resistant substrains are quantitatively correlated with the concentration of the specific chemical agent in the medium.
- Differences in light output due to the presence of other toxic or non- toxic agents are generally not significant and are not detected by the electro-optic sensor 15 because the sensitive substrain 12 and resistant " substrain 13 are derived from the same parent strain of photoluminescent microorganism, and they have the same sensitivity to other toxic or non-toxic agents.
- the measured light output from the electro- optic sensor 15 is conditioned and compared by the signal conditioning electronics 20.
- the signal conditioning electronics are a self-normalizing difference circuit.
- the self-normalizing difference circuit adjusts the readings of the electro-optic sen ⁇ sors 15 for the decrease of photoluminescence from the microorganisms due to the lapse of time, and can be used to adjust for temperature and humidity variations.
- the signal from the signal conditioning electronics 20 is recorded by the logic control electronics 21, which evaluate the signals to determine the presence of the specific chemical agent that caused the particular decrease in luminescence of the sensitive substrain 12.
- the logic control electronics 21 also can be programmed to indi ⁇ cate when a threshold concentration of a chemical agent is present so as to activate the alarm display elec ⁇ tronics 22.
- the logic control electronic control 21 also compiles the differences in photoluminescence between the sensitive and resistant substrains for two or more pairs of substrains in which each pair of sen- sitive and resistant substrains is derived from a parent strain based upon selection of mutants sensitive and resistant to the chemical agent, so as to distin ⁇ guish among a number of chemical agents.
- the alarm-display electronics 22 provides a visual reading of the various operational parameters and concentration levels. Various types of visual, auditory, and tactile alarms can be incorporated in the bioluminescent chemical system to warn of the presence of . toxic levels of a chemical agent.
- An environmental control module 23 monitors and controls the various external parameters to insure a reliable operation of the unit 11.
- all of the electronic cir ⁇ cuitry of the bioluminescent system, with the exception of the alarm-display electronics 22 and the electro- optic sensors 15, can be incorporated into a single hybrid chip.
- the parent strain of microorganism used in the mutant derivation of each pair of sensitive and resistant substrains may be any photoluminescent micro- organism such as bacteria or fungi.
- Bacteria such as Photobacteriu fischeri, Photobacterium phosphoreum, Photobacterium leioqnathi, and Vibrio harveyi may be employed.
- Bioluminescent fungi such as Armillarin mellea, Panus stipticus.
- Mycelium X, and members of the genus Mycena are also useful for this purpose.
- any photoluminescent organism manipulable through established microbial genetic methods, serves as a basis for the development of a specific detector system in which sensitive and resistant substrains are mutationally derived or derived by direct manipulation of genetic material.
- sub ⁇ strains of Photobacterium leioqnathi that are sensitive or resistant to acetone can be derived by two processes which differ only in the method of final selection of the substrain.
- a viable, actively grow ⁇ ing culture of the microorganism is exposed for defined periods of time to either nitrosoguanidine or ultra ⁇ violet light under conditions such that approximately 98 percent of the cells in the culture are killed.
- the remaining viable cells are rescued and shown to be a random assortment of mutant types, with the nature of genetic lesion a function of mutagen employed.
- single colony isolates of the mutant sub- strains are screened by the addition of acetone to the
- OMPI liquid cultures while a determination of the changes in light output of the microorganisms is made. This technique results in a pure culture isolation of ace ⁇ tone sensitive and resistant substrains. The same methodology can be used for the isolation of substrains sensitive or resistant to benzene.
- the ultra ⁇ violet mutated cultures of Photobacterium leioqnathi are designed to expose evenly a large number of bacter- ial colonies to a specific chemical agent. An immedi ⁇ ate comparison of luminescence of cultures is per ⁇ formed.
- air is pumped through a sparger submerged in acetone, and a petri dish containing the colonies of mutated bacteria are then exposed to the acetone saturated air.
- the petri dishes are illumi-*- nated from the side with a red light which makes lumi ⁇ nescent colonies appear blue and nonluminescent colo- ' nies appear red.
- the last colony to turn red is selected as a resistant sub ⁇ strain.
- This selection technique is used to derive sensitive and resistant substrains capable of detecting acetone at lower concentrations than substrains derived by the first methodology.
- the second screening technique can be used to derive sensitive and resistant substrains of Photobacterium phosphoreu capable of detecting chloroform or formaldehyde.
- the genetically derived and screened sub ⁇ strains can be preserved by lyophilization so that they may be stored until needed. As a first step in the lyophilization process, the microorganism cells are grown in the appropriate liquid medium until lumines- cence is maximum.
- the lyophilized cultures when activated with an"appropriate activation solution, emit light which is comparable to that observed before lyophilization. Depending upon the strain of microorganisms, the light emission continues for several hours and as long as 24 hours. Although the quantity of light decreases with time, the electronics of the detection system automati- cally adjusts to this decrease in luminescence. Accordingly, a single lyophilized packet of photolumi ⁇ nescent bacteria, after reconstitution with an activa ⁇ tion solution, could be used in the detection device of the present invention for periods of 24 hours even as the photoluminescence of the bacteria naturally decreases.
- the bioluminescent chemical system 10 was evaluated for its ability to detect and quantify
- the genetically selected substrains were lyo ⁇ philized by growing the cells until luminescence was maximum, and the cells were harvested by centrifuga ⁇ tion.
- the substrains were suspended in a skim milk cryoprotective solution. The suspensions were frozen, dried under a vacuum, and stored until needed. The cultures were protected from oxygen, water, and light.
- the cells were activated just prior to use. After activation, the activated cells were exposed, in an aerosol chamber, to defined concentrations of acetone or benzene.
- the benzene detection system was evaluated for its response to the structurally similar chemical toluene. The results of these procedures are summarized in the following table:
- ' "detection” refers to a concen ⁇ tration of chemical agent sufficient to be sensed as indicated by the lighting of the first light of the detector (approximately a 1% light level change).
- concentrations of the specific chemical agents increase, the signal generated by the light output dif ⁇ ference between the sensitive and resistant substrains increases monotonically. Therefore, quantification of response over a given concentration range of specific agents is straightforward.
- higher concen ⁇ trations of acetone and benzene would trigger the next optical indicators and the audible alarm system. The response of these functions was dependent upon the sen ⁇ sitive and resistant substrains employed and the sensi ⁇ tivity setting of the detection device.
- the luminescence from the microbial cells depends upon the maintenance of the cells in an optimal physiological state. An adequate supply of oxygen is required, and dehydration of the cultures should be avoided. Nutrients and an appropriate transparent matrix .material for support of the cells should be pro ⁇ vided.
- FIGS 2 and 2a illustrate a storage and activation system.
- the lyophilized cells are sealed in a clear, semipermeable plastic capsule 25.
- Each cap- _ sule 25 is divided into a microorganism compartment 26 and an activation solution compartment 27.
- the lyo ⁇ philized microorganisms are enclosed in the microorgan ⁇ ism compartment 26, and the activation solution is enclosed in the activation solution compartment 27.
- each compartment of capsule 25 is ruptured so that the activation solution in the activation solution compart ⁇ ment 27 enters the microorganism compartment 26 to activate the lyophilized microorganisms contained therein.
- a single membrane could be used to separate the microorganisms from the activation solution in capsule 25.
- activated microorganism mixture may be spread over the surface of a thin
- agar sorbent or hydrophilic polymer The spread of activated material occurs between the top 29 and bottom 30 layers formed from a transparent, semi- permeable membrane substance.
- the activated microorganisms can be spread over sorbent surface 31 which is not covered with a top layer 29. This latter technique would allow for the measurement of chemical agents which might not be capable of penetrating a semipermeable membrane.
- a chamber 32, enclosing a sorbent layer 31, is formed from the bottom layer 30 and the top layer 29 to allow the luminescent microorganisms to form a defined layer.
- oxygen, other gases, and solutes trapped in aerosols, along with the specific chemical agents to be detected, will pass through the polymer membranes 29 and 30 and readily contact the microorganism.
- the membrane pre ⁇ vents the premature desiccation of the culture without need for humidification, while preventing the direct contact of the microorganisms, media, or other liquids with the internal parts of the detection device.
- the lyophilized microorganisms Prior to activation, the lyophilized microorganisms remain viable and packaged in a potent state for a period of months to years.
- a plurality of capsules 25 can be formed in a sheet unit 35, as shown in Figure 2, with a sprocket 41 advancing the capsules 25 from a supply roll 42 to an expended roll 43 in a manner similar to the advancement of a film roll in a camera.
- the sheet unit 35 is pref ⁇ erably made of a clear, semipermeable plastic so that the capsules 25 can be easily formed therein.
- the storage and activation system shown in Figure 2 pro ⁇ vides a number of ready-to-activate sheet units 35 which can be located sequentially along a roll.
- a pack 37 including a plurality of detection units 38, having its own battery 40 and electronic system 39, can be used.
- a new detection unit 38 is automatically
- the battery pack 40 is capable of providing power sufficient for device operation over the lifetime of the photolumines ⁇ cence detection units 38 included in pack 37. This fail-safe feature alleviates concern about battery failure as a maintenance function separate from the microbial lifetime.
- the material which encapsulates the lyophil ⁇ ized microorganisms and solution in the microorganism compartment 26 and the activation solution compartment 27, respectively, is preferably made from a material, such as polylactate, which dissolves slowly and at a controlled rate.
- a material such as polylactate
- the microorganism cells are released and activated to exhibit lumines- cence.
- the rate of matrix dis ⁇ solution and cell replacement is equivalent to the rate of loss of cell luminescence.
- an electronic logic con ⁇ trol circuit 21 is utilized. For example, if one is interested in determining the presence or concentration of three agents A, B, and C, and if it was possible to develop microbial substrains sensitive or resistant to any two of the agents but not all three simultaneously (for example, abC is a strain that is resistant to agents A and B but sensitive to agent C), then three strains are needed, namely, abC, aBc, and Abe. In the following, small letters refer to resistance to the ' chemical agent and capital letters refer to sensitivity to the chemical agent. Using these three strains (abC, aBc, and Abe), the following truth table is generated:
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Abstract
Le système chimique bioluminescent décrit dans la présente invention est capable de détecter la présence d'un ou plusieurs agents chimiques dans un milieu liquide, de vapeur ou aérosol en utilisant des sous-souches de micro-organismes photoluminescents dérivées par mutation génétique à partir d'une souche parentale. Pour la détection d'un agent chimique spécifique, des sous-souches sensibles et résistantes sont sélectionnées génétiquement à partir de la même souche parentale de micro-organismes photoluminescents. L'émission lumineuse de la sous-souche sensible décroît en présence de l'agent chimique spécifique. Par opposition, l'émission ou puissance lumineuse de la sous-souche résistante ne change pas ou ne change pas parallèlement à celle de la sous-souche sensible, lorsqu'elle est exposée au même agent chimique. La différence relative de photoluminescence entre la sous-souche sensible et la sous-souche résistante est comparée de manière à indiquer la quantité ou la présence de l'agent chimique. De manière à permettre le stockage des sous-souches sélectionnées génétiquement pendant de longues périodes de temps entre le moment de la sélection génétique et le moment où elles sont finalement utilisées, les sous-souches sont lyophilisées. Immédiatement avant leur utilisation, les sous-souches sont activées en mélangeant les micro-organismes avec une solution d'activation de manière à leur redonner leur luminescence.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US52379483A | 1983-08-16 | 1983-08-16 | |
US523,794 | 1983-08-16 |
Publications (1)
Publication Number | Publication Date |
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WO1985000890A1 true WO1985000890A1 (fr) | 1985-02-28 |
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Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1984/001217 WO1985000890A1 (fr) | 1983-08-16 | 1984-07-30 | Systeme chimique bioluminescent et methode de detection de la presence d'agents chimiques dans un milieu |
Country Status (2)
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EP (1) | EP0153366A1 (fr) |
WO (1) | WO1985000890A1 (fr) |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2574430A1 (fr) * | 1984-12-06 | 1986-06-13 | Orgenics Ltd | Ensemble d'analyse microbiologique et procede de mise en evidence des inhibiteurs de la synthese biomoleculaire de novo |
FR2612295A1 (fr) * | 1987-03-11 | 1988-09-16 | Abensour David | Micro-analyseur automatique par colorimetrie ou bioluminescence |
EP0293775A2 (fr) * | 1987-06-05 | 1988-12-07 | Henkel Kommanditgesellschaft auf Aktien | Procédé pour déterminer l'action désinfectante d'un désinfectant et bande d'analyse pour cela |
DE3833628A1 (de) * | 1988-10-03 | 1990-04-12 | Genlux Forschungsgesellschaft | Verfahren zum nachweis und zur identifikation toxischer substanzen mit hilfe klonierter mikroorganismen |
DE3902982A1 (de) * | 1989-02-01 | 1990-08-02 | Genlux Forschungsgesellschaft | Verfahren zum nachweis von quecksilber mit hilfe von durch quecksilber zu erhoehter biolumineszenz angeregter mikroorganismen |
EP0527099A1 (fr) * | 1991-08-07 | 1993-02-10 | Gie Anjou-Recherche | Automate de détection de pollution en milieu aqueux mettant en oeuvre un test sur microorganisme. |
WO1993005142A1 (fr) * | 1991-09-11 | 1993-03-18 | Knight Scientific Limited | Appareil de contrôle de liquides |
FR2686350A1 (fr) * | 1992-01-17 | 1993-07-23 | Rochas | Methode d'evaluation de l'innocuite de produits au moyen de mesures de bioluminescence. |
EP0588139A1 (fr) * | 1992-09-10 | 1994-03-23 | Bayer Ag | Procédé analytique pour la recherche de constituants toxiques dans des mélanges |
WO1995010767A1 (fr) * | 1993-10-15 | 1995-04-20 | Merck Patent Gmbh | Procede de dosage |
DE4415866C1 (de) * | 1994-05-05 | 1995-06-22 | Draegerwerk Ag | Vorrichtung zum Nachweis von gas- und dampfförmigen Komponenten eines Gasgemisches |
US5827678A (en) * | 1993-09-08 | 1998-10-27 | Merck Patent Gmbh | Assay reagent comprising killed bacterial cells which retain functional metabolic activity |
US6017722A (en) * | 1991-04-04 | 2000-01-25 | Board Of Regents, The University Of Texas System | Luminous bacteria and methods for the isolation, identification and quantitation of toxicants |
US7256009B2 (en) | 1991-04-04 | 2007-08-14 | Becvar James E | Luminous bacteria and methods for the isolation, identification and quantitation of toxicants |
US7767447B2 (en) | 2007-06-21 | 2010-08-03 | Gen-Probe Incorporated | Instruments and methods for exposing a receptacle to multiple thermal zones |
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CHEMICAL ABSTRACTS, Vol. 100, No. 24, 11 June 1984 (Columbus, Ohio, US) F. KOESSLER: "Bacterial Luminescence - a Sensitive Tool Indicating Environmental Pollution", see page 687, Abstract No. 202521q, & Biolyumin. Tikhom Okeane, Mater. Simp. Tikhookean. Nauchn. Kongr., 14th 1979 (pub. 1982), pages 394 - 403 (Eng) * |
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Cited By (35)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR2574430A1 (fr) * | 1984-12-06 | 1986-06-13 | Orgenics Ltd | Ensemble d'analyse microbiologique et procede de mise en evidence des inhibiteurs de la synthese biomoleculaire de novo |
FR2612295A1 (fr) * | 1987-03-11 | 1988-09-16 | Abensour David | Micro-analyseur automatique par colorimetrie ou bioluminescence |
EP0293775A2 (fr) * | 1987-06-05 | 1988-12-07 | Henkel Kommanditgesellschaft auf Aktien | Procédé pour déterminer l'action désinfectante d'un désinfectant et bande d'analyse pour cela |
EP0293775A3 (fr) * | 1987-06-05 | 1989-05-31 | Henkel Kommanditgesellschaft auf Aktien | Procédé pour déterminer l'action désinfectante d'un désinfectant et bande d'analyse pour cela |
DE3833628A1 (de) * | 1988-10-03 | 1990-04-12 | Genlux Forschungsgesellschaft | Verfahren zum nachweis und zur identifikation toxischer substanzen mit hilfe klonierter mikroorganismen |
DE3902982A1 (de) * | 1989-02-01 | 1990-08-02 | Genlux Forschungsgesellschaft | Verfahren zum nachweis von quecksilber mit hilfe von durch quecksilber zu erhoehter biolumineszenz angeregter mikroorganismen |
US7713690B1 (en) | 1991-04-04 | 2010-05-11 | Becvar James E | Luminous bacteria and methods for the isolation, identification and quantitation of toxicants |
US6017722A (en) * | 1991-04-04 | 2000-01-25 | Board Of Regents, The University Of Texas System | Luminous bacteria and methods for the isolation, identification and quantitation of toxicants |
US7256009B2 (en) | 1991-04-04 | 2007-08-14 | Becvar James E | Luminous bacteria and methods for the isolation, identification and quantitation of toxicants |
EP0527099A1 (fr) * | 1991-08-07 | 1993-02-10 | Gie Anjou-Recherche | Automate de détection de pollution en milieu aqueux mettant en oeuvre un test sur microorganisme. |
FR2680247A1 (fr) * | 1991-08-07 | 1993-02-12 | Anjou Rech | Automate de detection de pollution en milieu aqueux mettant en óoeuvre un test sur microorganisme. |
WO1993005142A1 (fr) * | 1991-09-11 | 1993-03-18 | Knight Scientific Limited | Appareil de contrôle de liquides |
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