Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
• • 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
• • 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
  • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage

Definitions

• the present invention relates to a reagent, a measurement method, and a measurement device, and in particular to a reagent, a measurement method, and a measurement device that cause target microorganisms contained in a sample to fluoresce.
• testing has to rely on the culture method set by the government, which takes 48 hours and incurs huge costs. Companies that do not have their own testing system outsource all testing. However, these testing systems have a big problem. That is the time required for testing. Most foods are shipped immediately after production because freshness is important. In other words, even though the microbial test results for the products produced on the day can only be determined within 48 hours, the products are shipped and eaten before the test results are available. In the unlikely event that food poisoning occurs due to microbial contamination of food shipped earlier, the cause will not be known until later. This problem is a common theme worldwide. What is needed is to be able to ship products with peace of mind after the test results are available. In recent years, various simple test kits other than the culture method have been sold, but they take more than two hours and are simple, so there are problems with accuracy.
• the present invention was made in consideration of these circumstances, and aims to provide a reagent, a measurement method, and a measurement device that can measure microorganisms quickly and accurately.
• the reagent of the present invention is a reagent that causes a target microorganism contained in a specimen to fluoresce, and is characterized by selectively causing the target microorganism to fluoresce.
• the present invention by selectively making the target microorganisms fluorescent, it is possible to selectively measure the microorganisms, thereby enabling rapid and accurate measurement of the microorganisms.
• the target microorganism as E. coli and selectively making the E. coli fluorescent in the sample, it is possible to selectively measure E. coli in the sample, thereby enabling rapid and accurate measurement of E. coli.
• the bacteriophage t4 and/or bacteriophage t4 DNA is bound to a specific fluorescent reagent.
• E. coli which is a target microorganism contained in the sample
• bacteriophage t4 and/or bacteriophage t4 DNA bound to a specific fluorescent reagent thereby causing the E. coli to fluoresce.
• the specified fluorescent reagent is preferably FDA (fluorescein diacetate) and/or PI (propidium iodide).
• the measurement method of the present invention is a method for measuring target microorganisms contained in a sample using a reagent that causes the target microorganisms to fluoresce, and is characterized in that the target microorganisms are selectively made to fluoresce.
• the present invention by selectively making the target microorganisms fluorescent, it is possible to selectively measure the microorganisms in a sample, thereby enabling rapid and accurate measurement of microorganisms.
• the target microorganism is E. coli
• the reagent selectively causes the E. coli in the sample to fluoresce, allowing the E. coli to be selected and measured in the sample, thereby enabling rapid and accurate measurement of E. coli.
• the reagent is preferably a reagent in which a specific fluorescent reagent is bound to bacteriophage t4 and/or bacteriophage t4 DNA.
• the reagent is a reagent in which bacteriophage t4 and/or bacteriophage t4 DNA is combined with a specific fluorescent reagent, and it is preferable to selectively infect Escherichia coli, which is a target microorganism contained in the specimen, with the reagent to cause the Escherichia coli to fluoresce.
• the specified fluorescent reagent is preferably FDA (fluorescein diacetate) and/or PI (propidium iodide).
• the black filter impregnated with the reagent and the specimen containing the microorganism is placed in the measurement circle marked on the slide glass, and then covered with a cover glass to prepare a preparation.
• the measurement circles are marked on the glass slide in multiple places, and the black filter impregnated with a specimen containing the microorganism is placed in each of the multiple marked measurement circles, allowing multiple specimens containing microorganisms to be measured in parallel, improving measurement efficiency.
• the stage on which the preparation is placed can be moved along the X and Y axes to capture an image of the preparation.
• a CMOS sensor can be used to capture images of the slide, correct the captured images, and analyze the captured images.
• the tilt can be corrected even if the stage is tilted.
• the target microorganism is recognized from the captured image based on predetermined recognition parameters, and nothing other than the target microorganism is recognized, making it possible to efficiently recognize only the target microorganism.
• the recognition parameters may include at least three of the shape of the target microorganism, the size of the target microorganism, the fluorescence intensity of the target microorganism, and the shade of the fluorescence of the target microorganism.
• the measurement device of the present invention is a measurement device that measures target microorganisms contained in a sample using a reagent that causes the target microorganisms to fluoresce, and is characterized in that the target microorganisms are selectively made to fluoresce.
• the present invention by selectively making the target microorganisms fluorescent, it is possible to selectively measure the microorganisms, thereby enabling rapid and accurate measurement of the microorganisms.
• the target microorganism is E. coli
• the reagent selectively causes the E. coli in the sample to fluoresce, allowing the E. coli to be selected and measured in the sample, thereby enabling rapid and accurate measurement of E. coli.
• the reagent is preferably a reagent in which a specific fluorescent reagent is bound to bacteriophage t4 and/or bacteriophage t4 DNA.
• the reagent is a reagent in which bacteriophage t4 and/or bacteriophage t4 DNA is combined with a specific fluorescent reagent, and it is preferable to selectively infect Escherichia coli, which is a target microorganism contained in the specimen, with the reagent to cause the Escherichia coli to fluoresce.
• the specified fluorescent reagent is preferably FDA (fluorescein diacetate) and/or PI (propidium iodide).
• the black filter impregnated with the reagent and the specimen containing the microorganism is placed in the measurement circle marked on the slide glass, and then covered with a cover glass to prepare a preparation.
• the measurement circles are marked on the glass slide in multiple places, and the black filter impregnated with a specimen containing the microorganism is placed in each of the multiple marked measurement circles, allowing multiple specimens containing microorganisms to be measured in parallel, improving measurement efficiency.
• the stage on which the preparation is placed can be moved along the X and Y axes to capture an image of the preparation.
• a CMOS sensor can be used to capture images of the slide, correct the captured images, and analyze the captured images.
• the tilt can be corrected even if the stage is tilted.
• the target microorganism is recognized from the captured image based on predetermined recognition parameters, and nothing other than the target microorganism is recognized, making it possible to efficiently recognize only the target microorganism.
• the recognition parameters may include at least three of the shape of the target microorganism, the size of the target microorganism, the fluorescence intensity of the target microorganism, and the shade of the fluorescence of the target microorganism.
• the present invention aims to provide a reagent, a measurement method, and a measurement device that can measure microorganisms quickly and accurately.
• FIG. 2 is a diagram showing a configuration of a reagent according to an embodiment of the present invention.
• FIG. 13 shows the state in which the reagent is taken up by E. coli.
• 1A to 1C are diagrams showing a method for preparing a slide used in a measuring method and a measuring device according to an embodiment of the present invention.
• 4 is a diagram continuing from FIG. 3 showing a method for preparing a slide used in the measuring method and measuring device according to an embodiment of the present invention.
• FIG. 5 is a view following FIG. 4 showing a method for preparing a slide used in the measuring method and measuring device according to an embodiment of the present invention.
• 5A to 5C are diagrams showing a method for preparing a slide used in the measuring method and measuring device according to an embodiment of the present invention.
• 7 is a view following FIG. 6 showing a method for preparing a slide used in the measuring method and measuring device according to an embodiment of the present invention.
• 8 is a view following FIG. 7 showing a method for preparing a slide used in the measuring method and measuring device according to an embodiment of the present invention.
• 2 is a diagram showing a configuration of a microscope device used in the measurement method and measurement device.
• FIG. 13 is another diagram showing the configuration of the microscope apparatus used in the measurement method and measurement apparatus.
• FIG. 1 is a diagram showing the configuration of a reagent according to an embodiment of the present invention
• FIG. 2 is a diagram showing the state in which the reagent has been taken up into E. coli.
• the reagent 1 is a reagent in which [bacteriophage t4 and/or bacteriophage t4 DNA] 1A and a specified fluorescent reagent 1B are mixed and combined in advance.
• Bacteriophage T4 is a species of bacteriophage that infects Escherichia coli 102 and is classified as a double-stranded DNA virus.
• Bacteriophage t4 DNA is the DNA of bacteriophage T4, and can be obtained, for example, by extracting DNA from bacteriophage T4.
• the reagent 1 is a reagent 1 that causes the target microorganism 101 contained in the specimen 100 shown in FIG. 2 to fluoresce, and can selectively cause the target microorganism 101 to fluoresce.
• the target microorganism 101 is E. coli 102, and the E. coli 102 can be selectively caused to fluoresce in the specimen 100.
• the reagent 1 is a reagent in which [bacteriophage t4 and/or bacteriophage t4 DNA] 1A and a specific fluorescent reagent 1B are mixed and combined in advance, and the reagent 1 is selectively infected with E. coli 102, which is the target microorganism 101 contained in the specimen 100, and taken up by the E. coli 102, thereby making the E. coli 102 fluorescent.
• the specific fluorescent reagent 1B is a DNA fluorescent reagent, and can be FDA (fluorescein diacetate) and/or PI (propidium iodide).
• FDA fluorescein diacetate
• PI propidium iodide
• the reagent 1 can selectively cause the target microorganisms 101 to fluoresce, allowing the microorganisms 101 to be selected and measured, thereby enabling rapid and accurate measurement of the microorganisms 101.
• the target microorganism 101 as E. coli 102 and selectively making the E. coli 102 fluorescent in the specimen 100, it is possible to selectively measure the E. coli 102 in the specimen 100, thereby enabling rapid and accurate measurement of the E. coli 102.
• FIGS. 9 and 10 are diagrams showing the configuration of a microscope apparatus used in the same measurement method and measurement apparatus.
• the above-mentioned reagent 1 is used in the measurement method and measurement device 200 according to the embodiment of the present invention. That is, the measurement method and measurement device 200 are a measurement method and measurement device 200 that measure target microorganisms 101 contained in a specimen 100 using a reagent 1 that causes the target microorganisms 101 to fluoresce, and can selectively cause the target microorganisms 101 to fluoresce.
• the measurement device 200 has a fluorescence processing device 60 and a microscope device 70, which will be described later.
• the target microorganism 101 is E. coli 102
• the reagent 1 can selectively cause the E. coli 102 in the sample to fluoresce.
• Reagent 1 is a reagent in which [bacteriophage t4 and/or bacteriophage t4 DNA] 1A is premixed and bound to a specific fluorescent reagent 1B.
• Bacteriophage T4 is a species of bacteriophage that infects E. coli 102 and is classified as a double-stranded DNA virus.
• Bacteriophage t4 DNA is the DNA of bacteriophage T4, and can be obtained, for example, by extracting DNA from bacteriophage T4.
• the reagent 1 is a reagent 1 that causes the target microorganism 101 contained in the specimen 100 shown in FIG. 2 to fluoresce, and can selectively cause the target microorganism 101 to fluoresce.
• the target microorganism 101 is E. coli 102, and the E. coli 102 can be selectively caused to fluoresce in the specimen 100.
• the reagent 1 is a reagent in which [bacteriophage t4 and/or bacteriophage t4 DNA] 1A and a specific fluorescent reagent 1B are mixed and combined in advance, and the reagent 1 is selectively infected with E. coli 102, which is the target microorganism 101 contained in the specimen 100, and taken up by the E. coli 102, thereby making the E. coli 102 fluorescent.
• the specific fluorescent reagent 1B is a DNA fluorescent reagent, and can be FDA (fluorescein diacetate) and/or PI (propidium iodide).
• FDA fluorescein diacetate
• PI propidium iodide
• E. coli 102 can be measured using a specified preparation 10.
• the method for preparing the preparation 10 is described as follows.
• a black filter 20 is prepared.
• a black filter 20 is impregnated with a reagent 1 that selectively causes the E. coli 102 to fluoresce, and the black filter 20 impregnated with the reagent 1 can be used to perform a process that selectively causes the E. coli 102 to fluoresce.
• a black filter 20 is impregnated with a reagent 1 that selectively causes E. coli 102 to fluoresce
• the black filter 20 impregnated with the reagent 1 is impregnated with a specimen 100 that contains E. coli 102, making it possible to perform a process that selectively causes E. coli 102 to fluoresce.
• a black filter 20 impregnated with a specimen 100 containing reagent 1 and E. coli 102 is placed within the marked measurement circle 40, and then, as shown in Figure 8, it is covered with a cover glass 50 to prepare a preparation 10.
• Multiple measurement circles 40 are marked on the glass slide 30, and a black filter 20 impregnated with a specimen 100 containing E. coli 102 is placed in each of the multiple marked measurement circles 40.
• the prepared preparation 10 can be used to perform fluorescence processing by a fluorescence processing device 60, as shown in Figures 9 and 10, and images can be captured, corrected, and analyzed by a microscope device 70.
• the microscope device 70 is a fluorescence microscope device.
• the fluorescence processing device 60 has an excitation light irradiation unit 61 and a wavelength measurement unit 62.
• the excitation light irradiation unit 61 can irradiate the preparation 10 (more specifically, the multiple black filters 20 impregnated with the reagent 1 and the E. coli 102) with excitation light of a predetermined wavelength.
• the wavelength measurement unit 62 can measure the wavelength of the fluorescence from the preparation 10 (more specifically, the multiple black filters 20 impregnated with the reagent 1 and the E. coli 102) that has absorbed the excitation light.
• the microscope device 70 has a stage 71 that moves along the XY axis. That is, in the microscope device 70, the stage 71 on which the preparation 10 is placed can be moved along the XY axis to capture multiple images of the preparation 10 that has absorbed the excitation light (more specifically, multiple black filters 20 impregnated with the reagent 1 and E. coli 102).
• the microscope device 70 further includes a memory unit 72, an imaging unit 73, a correction unit 75, and an analysis unit 74.
• the memory unit 72 can store data on the standard shape of the target microorganism 101, E. coli 102, the standard size of the target microorganism 101, E. coli 102, the standard fluorescence intensity of the target microorganism 101, E. coli 102, and the standard fluorescence shading of the target microorganism 101, E. coli 102.
• the photographing unit 73 photographs multiple images of the preparation 10 using a CMOS sensor for each of the multiple black filters 20, the correction unit 75 corrects the images photographed by the photographing unit 73, and the analysis unit 74 can qualitatively and/or quantitatively analyze the images photographed by the photographing unit 73.
• the correction unit 75 can correct the image captured by the imaging unit 73 so as to correct differences in thickness and/or shape between the glass slides 30 in the preparation 10.
• the correction unit 75 can correct the image captured by the photographing unit 73 so as to correct the difference between the thickness and/or shape of the slide glass 30 to be photographed and the thickness and/or shape of the standard slide glass 30.
• the correction unit 75 can also correct the image captured by the photographing unit 73 so that the inclination of the stage 71 is corrected. That is, the correction unit 75 can correct the image captured by the photographing unit 73 so that the inclination of the stage 71 with respect to a predetermined direction, more specifically, the horizontal direction, is corrected.
• the photographing unit 73 can also capture the image while enlarging or reducing the image by changing the distance from the preparation 10 multiple times.
• the photographing unit 73 has a function of measuring the surface of the specimen 100 at a distance of three or more measurement points for each measurement of the thickness and individual differences in the shape of the slide glass 30 and the inclination of the stage 71 on which the slide glass 30 is placed, and capturing the image while correcting the individual differences (thickness and distortion) and inclination of the slide glass 30 with the zoom of the photographing unit 73 or the correction unit 75.
• the analysis unit 74 can recognize the target microorganism 101, E. coli 102, from the image captured by the imaging unit 73 based on predetermined recognition parameters, and can perform analysis so as not to recognize anything other than the target microorganism 101, E. coli 102.
• the recognition parameters can include, for example, at least three of the shape of the target microorganism 101, E. coli 102, the size of the target microorganism 101, E. coli 102, the intensity of the fluorescence of the target microorganism 101, E. coli 102, and the shade of the fluorescence of the target microorganism 101, E. coli 102.
• the analysis unit 74 reads out data on the standard shape of the target microorganism 101, E. coli 102, the standard size of the target microorganism 101, the standard fluorescence intensity of the target microorganism 101, and the standard fluorescence shading of the target microorganism 101, and compares the read out data with the image captured by the image capture unit 73 (more specifically, the image corrected by the correction unit 75) to analyze whether the captured image (more specifically, the corrected image) matches at least three of the standard shape of the target microorganism 101, E. coli 102, the standard size of the target microorganism 101, the standard fluorescence intensity of the target microorganism 101, and the standard fluorescence shading of the target microorganism 101, within a predetermined range.
• the analysis unit 74 can consider that the microorganism 101 in the captured image (more specifically, the corrected image) is E. coli 102, which is the target microorganism 101.
• the microorganism 101 in the captured image can be considered to be the target microorganism 101, E. coli 102, when it matches within a specified range at least any three of the standard shape of the target microorganism 101, E. coli 102, the standard size of the target microorganism 101, E. coli 102, the standard fluorescence intensity of the target microorganism 101, E. coli 102, and the standard fluorescence shade of the target microorganism 101, E. coli 102.
• the specimen 100 may contain food residue, dirt, and other foreign matter, and even if fluorescent staining is performed as described above, it may be difficult to distinguish between microorganisms 101 and non-microorganisms.
• recognition parameters as described above, it is possible to measure only microorganisms 101 such as E. coli 102 and not measure anything other than microorganisms 101.
• the shape of the target microorganism 101, E. coli 102, the size of the target microorganism 101, the fluorescence intensity of the target microorganism 101, and the fluorescence shade of the target microorganism 101, E. coli 102 are affected by differences in thickness and/or shape between the glass slides 30 in the preparation 10 and the inclination of the stage 71, causing the image to become blurred due to focus shifting.
• the correction unit 75 can correct the image captured by the image capture unit 73 so as to correct the differences in thickness and/or shape between the glass slides 30 in the preparation 10, and can correct the image captured by the image capture unit 73 so as to correct the inclination of the stage 71, so that such influences can be appropriately corrected to perform accurate image processing.
• the measurement method and measurement device 200 of the present invention can selectively cause the target microorganisms 101 to fluoresce, thereby enabling the microorganisms 101 to be selected and measured in the specimen 100, thereby enabling rapid and accurate measurement of the microorganisms 101.
• the target microorganism 101 is E. coli 102
• the reagent 1 selectively causes the E. coli 102 in the specimen 100 to fluoresce, thereby allowing the E. coli 102 to be selected and measured in the specimen 100, thereby enabling rapid and accurate measurement of the E. coli 102.
• the black filter 20 by impregnating the black filter 20 with a reagent 1 that selectively causes the E. coli 102 to fluoresce, and then performing a process to selectively cause the E. coli 102 to fluoresce using the black filter 20 impregnated with the reagent 1, the fluorescence of the E. coli 102 stands out against the black background, allowing the E. coli 102 to be measured quickly and accurately.
• multiple measurement circles 40 are marked on the glass slide 30, and a black filter 20 impregnated with a specimen 100 containing E. coli 102 is placed in each of the multiple marked measurement circles 40, so that multiple specimens 100 containing E. coli 102 can be measured in parallel, improving the efficiency of the measurement.
• the correction unit 75 can correct the image captured by the photographing unit 73 so that the differences in thickness and/or shape between the glass slides 30 in the preparation 10 are corrected, thereby correcting the differences even if the thickness and/or shape between the glass slides 30 differs.
• the correction unit 75 can correct the image captured by the image capture unit 73 so that the tilt of the stage 71 is corrected, thereby correcting the tilt even if the stage 71 is tilted.
• the correction unit 75 recognizes the target microorganism 101 from the image captured by the imaging unit 73 based on predetermined recognition parameters, and does not recognize anything other than the target microorganism 101, thereby enabling efficient recognition of only the target microorganism 101.
• the microorganism 101 is E. coli 102, but it is possible to appropriately accommodate a microorganism 101 other than E. coli 102.
• the black filter 20 is impregnated with the reagent 1 that selectively causes the E. coli 102 to fluoresce, and the black filter 20 impregnated with the reagent 1 is used to perform a process to selectively cause the E. coli 102 to fluoresce.
• the desired effect can also be achieved by impregnating the black filter 20 with the reagent 1 that selectively causes the microorganisms 101 other than the E. coli 102 to fluoresce, and using the black filter 20 impregnated with the reagent 1 to perform a process to selectively cause the microorganisms 101 other than the E. coli 102 to fluoresce.
• a black filter 20 impregnated with a specimen 100 containing reagent 1 and E. coli 102 is placed in the measurement circle 40 marked on the slide glass 30, and then covered with a cover glass 50 to prepare the preparation 10.
• the desired effect can also be achieved by placing a black filter 20 impregnated with a specimen 100 containing reagent 1 and a microorganism 101 other than E. coli 102 in the measurement circle 40 marked on the slide glass 30 and covering it with a cover glass 50 to prepare the preparation 10.
• the target microorganism 101, E. coli 102 is recognized from the image captured by the image capture unit 73 based on predetermined recognition parameters, and nothing other than the target microorganism 101, E. coli 102, is recognized.
• the recognition parameters include at least three of the shape of the target microorganism 101, E. coli 102, the size of the target microorganism 101, the fluorescence intensity of the target microorganism 101, E. coli 102, and the fluorescence shade of the target microorganism 101, E. coli 102.
• the desired effect can be achieved even if the target microorganism 101 is a microorganism 101 other than E. coli 102.
• the desired effect can be achieved by simply recognizing the target microorganism 101 from the image captured by the imaging unit 73 based on predetermined recognition parameters, and not recognizing anything other than the target microorganism 101, and the recognition parameters can include at least three of the shape of the target microorganism 101, the size of the target microorganism 101, the intensity of the fluorescence of the target microorganism 101, and the shade of the fluorescence of the target microorganism 101.
• a black filter 20 impregnated with a specimen 100 containing E. coli 102 is placed in each of the multiple marked measurement circles 40, but a black filter 20 impregnated with a specimen 100 containing E. coli 102 and a black filter 20 impregnated with a specimen 100 containing a microorganism 101 other than E. coli 102 may be placed in separate measurement circles 40 to simultaneously measure two or more types of microorganisms 101 (for example, to simultaneously measure the general bacterial count and E. coli count).
• Reagent 1A Bacteriophage t4 and/or bacteriophage t4 DNA 1B: Fluorescent reagent 10: Preparation 20: Filter 30: Slide glass 40: Measurement circle 50: Cover glass 60: Fluorescence treatment device 61: Excitation light irradiation unit 62: Wavelength measurement unit 70: Microscope device 71: Stage 72: Memory unit 73: Photography unit 74: Analysis unit 75: Correction unit 100: Sample 101: Microorganism 102: Escherichia coli 200: Measurement device

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• 2022
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