WO2014054494A1 - 微生物培養器、微生物検査キット、透析液の検査方法、微生物の培養方法、微生物の検査方法、及び微生物培養器の製造方法 - Google Patents
微生物培養器、微生物検査キット、透析液の検査方法、微生物の培養方法、微生物の検査方法、及び微生物培養器の製造方法 Download PDFInfo
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- WO2014054494A1 WO2014054494A1 PCT/JP2013/076022 JP2013076022W WO2014054494A1 WO 2014054494 A1 WO2014054494 A1 WO 2014054494A1 JP 2013076022 W JP2013076022 W JP 2013076022W WO 2014054494 A1 WO2014054494 A1 WO 2014054494A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/14—Bags
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/46—Means for fastening
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/04—Seals
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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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
- C12Q1/04—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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T29/00—Metal working
- Y10T29/49—Method of mechanical manufacture
- Y10T29/49826—Assembling or joining
Definitions
- This application discloses a microorganism incubator, a microorganism inspection kit, a dialysate inspection method, a microorganism culture method, a microorganism inspection method, and a microorganism incubator manufacturing method.
- Microorganisms are cultured using various media.
- the medium include a liquid medium and an agar medium that have been generally used.
- the liquid culture medium has no limitation on the amount of sample, but has a drawback that it is not quantitative and bulky.
- the upper limit of the amount of the sample solution in a petri dish having a diameter of 90 mm that is normally used for an agar medium is usually 1 mL, and it is difficult to inspect the sample solution exceeding 1 mL. Therefore, when inspecting a sample having a relatively low microorganism concentration, in order to inspect with a sample solution of about 1 mL, first, the original sample is subjected to enrichment culture and then subjected to the inspection.
- a sensitivity of 1 cfu / 25 g is required for the inspection of food poisoning bacteria such as Salmonella.
- dialysis water dialysis water
- a large amount of samples may be required for testing.
- dialysis water is required to have a high sensitivity capable of detecting less than one microorganism in 1 mL of a sample. If a sample solution exceeding 1 mL, for example 10 to 100 mL, can be directly and quantitatively tested for microorganisms. Simple and desirable.
- a microorganism incubator containing a sheet-like medium in a bag-like container is, for example, a sterilized microorganism incubator by opening a zipper at the opening of the bag-like container and placing it in an atmosphere of sterilization gas. be able to.
- the microorganism incubator sterilized in this way may have foreign substances such as microorganisms enter the container through the opening until the opening is closed after the sterilization.
- the bag-like container using zippers as the opening / closing material of the opening portion has low hermeticity, foreign substances may enter even when the opening portion is closed. It is difficult to put a large amount of a sample having a small number of bacteria in order to detect bacteria.
- the present application provides a microorganism culture device, a microorganism test kit, a dialysate test method, a microorganism culture method, a microorganism test method, and a microorganism culture that can suppress the presence of invading foreign substances and can contain a large amount of sample. It aims at providing the manufacturing method of a vessel.
- the present invention has provided an inlet for putting a sample in a culture bag separately from an opening portion for putting a medium inside.
- the present invention is a microbial incubator including a culture bag with an inlet filled with a sheet-like culture medium and sealed.
- a microbial incubator including a culture bag with an inlet filled with a sheet-like culture medium and sealed.
- a sample when a sample is placed in a culture bag, it is inserted through an injection port through which foreign matter is difficult to pass compared to an opened portion where the medium can be taken in and out. It is suppressed.
- the container is highly sealed and difficult to leak out, a large amount of sample can be placed.
- the inlet of the culture section may be any one that can be sealed after the sample has been injected.
- spout screw cap, push-in cap, crown, rubber plug, urethane plug Any one or more of a silicon stopper, a cork stopper, a Molton stopper, a cotton stopper, a paper stopper, a three-way cock, a fastener, a chuck and a zipper can be applied.
- the microorganism culture device may include a culture bag with an inlet sealed with a sheet-like culture medium inside, and a sterilization bag with the culture bag sealed inside. .
- a microbial incubator since the sterilization state outside the culture bag is maintained by the sterilization bag, there is a low possibility that a foreign substance will enter the culture bag when the sample is put into the culture bag.
- the microorganism culture vessel is for detecting aerobic bacteria
- the culture bag may be formed of an oxygen permeable material. If the culture bag is formed of an oxygen permeable material, oxygen is supplied from the outside to the inside of the culture bag, so that aerobic bacteria can be grown.
- the microorganism culture vessel is for detecting anaerobic bacteria
- the culture bag may be formed of an oxygen-impermeable material. If the culture bag is made of an oxygen-impermeable material, oxygen is not supplied from the outside to the inside of the culture bag, so that anaerobic bacteria can be grown.
- the microorganism incubator is for detecting microorganisms contained in the dialysate or reverse osmosis water
- the culture medium in the culture bag is a predetermined supplement for supplementing the nutrient components contained in the dialysate or reverse osmosis water. It may have a nutritional component. If the medium has a predetermined supplemental nutrient component that supplements the nutrient component originally contained in the dialysate, it is grown in a culture bag without killing microorganisms in the dialysate due to overnutrition. It is possible.
- the predetermined supplementary nutrients contained in the culture medium in the culture bag are, for example, 0.1 to 3.0 g of peptone, 0.1 to 3.0 g of yeast extract, and 0.1 to 0 per square meter.
- 0.5 g casein degradation product 0-0.5 g glucose, 0.075-0.3 g sodium pyruvate, 0-0.3 g dipotassium phosphate, 0.1-0.5 g It may be soluble starch, 0.01-0.05 g magnesium sulfate, 0.015-0.1 g tetrazolium salt or 0.15-1.0 g indoxyl derivative. If such a component is contained in the culture medium, microorganisms in the dialysate or reverse osmosis water can be grown in the culture bag.
- the predetermined supplementary nutrients contained in the culture medium in the culture bag are, for example, 0.38 to 9.0 g of meat extract or fish extract, 0.63 to 15.0 g of tryptone, 0 to It may be 3.0 g glucose, 0-0.3 g dipotassium phosphate, 0.015-0.1 g tetrazolium salt or 0.15-1.0 g indoxyl derivative. If such a component is contained in the culture medium, microorganisms in the dialysate or reverse osmosis water can be grown in the culture bag.
- the predetermined supplementary nutrients contained in the culture medium in the culture bag are, for example, 2.5 to 40.0 g of malt extract, 0 to 40.0 g of glucose, and 0.25 to 4.0 g per square meter.
- the present invention can also be understood from the aspect as a microorganism testing kit including any one of the above microorganism incubators.
- the present invention can be understood from the aspect as a method for inspecting dialysate.
- the present invention is sealed in a sterilization bag of a microorganism incubator provided with a culture bag with an inlet filled with a sheet-like culture medium and sealed, and a sterilization bag with the culture bag sealed inside.
- It may be a dialysate testing method in which microorganisms are cultured and the presence or amount of the cultured microorganisms is tested.
- the present invention can be grasped from the aspect as a microorganism culturing method.
- the present invention is sealed in a sterilization bag of a microorganism incubator provided with a culture bag with an inlet filled with a sheet-like culture medium and sealed, and a sterilization bag with the culture bag sealed inside.
- a sterilization bag of a microorganism incubator provided with a culture bag with an inlet filled with a sheet-like culture medium and sealed, and a sterilization bag with the culture bag sealed inside.
- Remove the stopped culture bag from the sterile bag open the culture bag inlet, put the sample solution into the culture bag from the inlet, seal the inlet and place the culture bag under microbial culture conditions. It may be a method for culturing microorganisms.
- the present invention can be grasped from the aspect as a microbe inspection method.
- the present invention may be a microorganism testing method for testing the presence or amount of microorganisms cultured by the microorganism cultivation method.
- the present invention can be understood from the aspect as a method for producing a microorganism incubator.
- the present invention is a method for producing a microorganism incubator comprising a culture bag with an inlet filled with a sheet-like culture medium and sealed, and a sterilization bag sealed with the culture bag inside.
- the culture bag in which the culture medium is put inside from the opened part of the culture bag is housed in a gas-permeable sterilization bag, the sterilization bag is sealed, and the sealed sterilization bag is placed in a sterilized gas atmosphere. Sterilize the medium in the culture bag contained in the sterilization bag, and close the opened part of the culture bag from the outside of the sterilization bag with the sterilization bag containing the culture bag sealed.
- the culture bag containing the sterilized medium may be sealed.
- a bag using a small opening portion such as a cap having a high sealing property as an injection port
- a sterilized microorganism incubator is prepared by storing a sheet-shaped medium in a sterilized container and placing it in a sterilized gas atmosphere, the size of the opening is small. Is likely to remain. If sterilization gas remains in the bag-like container, it affects the growth of microorganisms to be cultured. It is difficult to completely remove the residual gas from the container even if a gas-permeable material is used for the bag-like container.
- the microorganism incubator is manufactured by such a method, the culture bag is kept in the sterile bag until the culture bag is sealed after the medium is sterilized. Therefore, the possibility that foreign matter enters the culture bag after sterilization is extremely low.
- the microorganism incubator is manufactured by such a method, the culture bag remains in the sterilization bag until the culture bag is sealed after the medium is sterilized. The open part of the culture bag can be enlarged, and the sterilization gas remaining in the culture bag can be attracted and removed from the outside of the sterilization bag through the sterilization bag having gas permeability.
- the method for producing a microorganism incubator may be such that after the sterilization process and before the sealing process, the sterilized bag subjected to the sterilization process is placed in a vacuum to further remove the sterilization gas remaining in the sterilization bag. Good. If the removal treatment is further performed, it is possible to produce a microorganism incubator in which residual sterilization gas is suppressed.
- the method for manufacturing the microorganism culture device may be a method in which the culture bag is further taken out from the sterilization bag. If the removal process is further performed, it is possible to distribute the sterilized culture bags.
- the inner surface of the culture bag is made of a material that can be heat-welded at least at a lower temperature than the material that forms the inner surface of the sterilization bag among the materials that form the sterilization bag.
- the culture bag may be heated in a state of being closed from the outside to seal the culture bag. If the sealing process is performed by heat welding from the outside of the sterilization bag, the sealing process is easy, and the sealing property of the culture bag can be improved and the entry of foreign substances can be more reliably suppressed.
- FIG. 6 is an example of a cross-sectional view of a sterilization bag when cut along a line indicated by AA in FIG. 5. It is an example of the figure which showed a mode that the culture bag containing the culture medium was sealed.
- FIG. 9 is an example of a cross-sectional view of a sterilization bag and a culture bag when cut along the line indicated by reference numeral BB in FIG. 8. It is an example of the figure which showed a mode that the culture bag was taken out from a sterilization bag. It is an example of the figure which showed a mode that the sample was put in a culture bag. It is an example of the figure which showed the culture bag which concerns on a 1st modification. It is an example of the figure which showed the culture bag which concerns on a 2nd modification. It is an example of the figure which showed the culture bag which concerns on a 3rd modification.
- FIG. 1 is an example of an external view of a microorganism incubator according to an embodiment.
- the microorganism incubator 1 is a microorganism incubator in which a sheet-shaped medium 20 is put in a bag-shaped culture bag 10 and is accommodated in a sterilization bag 30.
- the microorganism culture device 1 is not limited to the one provided with the sterilization bag 30 that accommodates the culture bag 10, and the sterilization bag 30 may be omitted.
- the microbial incubator 1 may constitute all or part of a microbial test kit appropriately including convenient items when testing bacteria.
- the culture bag 10 is provided with an inlet 11 for injecting a sample into the container.
- the culture bag 10 is formed by welding the edges of two sheets of sheet-like material (for example, a heat-meltable material such as Polymer), and the edge portions 12A and 12B are welded as shown in FIG. Has been.
- the sterilization bag 30 is formed by welding the edges of two stacked sheets of material, and the edge portions 31A and 31B are welded as shown in FIG.
- the sample does not leak when the sample is added, is liquid impervious, and has oxygen permeability that allows aerobic bacteria to grow. Further, it is preferably transparent or translucent in order to observe the growth of microorganisms.
- oxygen permeability is 3mL / m 2/24 hours or more.
- a resin film such as Polymer as described above can be used.
- the thickness of the film at that time is suitably in the range of 150 ⁇ m.
- the resin include polyethylene, polypropylene, and composites thereof. For example, when using polypropylene, the thickness is desirably 150 ⁇ m or less from the viewpoint of gas permeability.
- the shape of the culture bag 10 is not particularly limited. For example, a square such as a square or a rectangle is convenient for mass production.
- the size of the culture bag 10 can be determined freely according to the amount of the sample. For example, if the assumed amount of the sample is 10 mL, a culture bag of about 100 ⁇ 120 mm to 160 ⁇ 200 mm is prepared. This is convenient.
- the injection port 11 of the culture bag 10 may be any as long as the sample can be injected into the container, and in addition to caps having excellent sealing properties, for example, zippers may be used.
- the microorganism incubator 1 can be manufactured, for example, by the following manufacturing process.
- FIG. 2 is an example of a diagram showing a state in which the culture medium 20 is put into the culture bag 10.
- the culture medium 20 is put into the culture bag 10 from the open portion 13 before the edge portion 12B is welded.
- FIG. 3 is an example of a diagram illustrating a state in which the culture bag 10 in which the medium 20 is placed is accommodated in the sterilization bag 30. After the culture medium 20 is put in the culture bag 10, the culture bag 10 with the open portion 13 open is put into the sterile bag 30 from the open portion 32 before the edge portion 31B is welded.
- FIG. 4 is an example of a diagram showing a state in which the sterilization bag 30 containing the culture bag 10 is sealed. After the culture bag 10 is accommodated in the sterilization bag 30, the open portion 32 is closed with the welding sealer 2 and the sterilization bag 30 is sealed. Thereby, the sterilization bag 30 will be in the state by which two sheet-like raw materials which comprise the sterilization bag 30 mutually weld by the edge part 31B shown in FIG. 1, and the sterilization bag 30 was sealed.
- FIG. 5 is an example of a diagram showing the sterilization bag 30 after the open portion 32 is closed with the welding sealer 2.
- the inside of the sterilization bag 30 is closed over the entire circumference by the edge portions 31A and 31B. Accordingly, the culture bag 10 in the sterilization bag 30 is isolated from microorganisms and other foreign matters outside the sterilization bag 30.
- FIG. 6 is an example of a cross-sectional view of the sterilization bag 30 taken along the line AA in FIG.
- the sterilization bag 30 is formed of one of the two sheet-like materials constituting the sterilization bag 30 with a gas permeable sheet 31, and the other can be fused to the sheet 31. It is made of a heat-meltable film 33. Since the sterilization bag 30 is configured as described above, the open portion 32 can be closed by the welding sealer 2, and the sterilization bag 30 can be placed in a predetermined gas atmosphere even when the open portion 13 is closed. For example, the predetermined gas permeates the sheet 31, and the contents in the sterilization bag 30 can be exposed to the predetermined gas.
- the sterilization bag 30 that has been sealed by closing the open portion 32 with the welding sealer 2 is placed in an atmosphere of sterilization gas, and the outside of the culture bag 10 accommodated in the sterilization bag 30.
- the medium 20 and the like in the culture bag 10 are sterilized.
- the sterilization bag 30 By placing the sterilization bag 30 in an atmosphere of sterilization gas, microorganisms attached to the outside and inside of the culture bag 10 accommodated in the sterilization bag 30, the medium 20 in the culture bag 10, etc. are killed.
- the gas permeable sheet 31 of the sterilization bag 30 is provided with indicators such as ink that changes color when the gas comes into contact with the sterilization bag 30, whether or not the microorganism incubator 1 has been sterilized. Discrimination is easy.
- the sterilization treatment include the following treatments. That is, for example, indicators that change color when in contact with gas are attached to an appropriate location such as the culture bag 10 or the sterilization bag 30, and the sterilization bag 30 containing the culture bag 10 is inserted into a sterilizer.
- the temperature setting of the sterilizer is, for example, about 45 ° C.
- the door of the sterilizer is closed and the inside of the can is depressurized (for example, a gauge pressure of about ⁇ 0.085 MPa) to make a vacuum state.
- EO (ethylene oxide) gas is introduced up to a predetermined pressure (for example, a gauge pressure of 0.110 MPa).
- a predetermined time for example, 12 hours after reaching the set pressure
- the sterilization action gradually proceeds in the sterilizer.
- the inside of the sterilizer is degassed and evacuated. Since the edge portion 12B of the culture bag 10 is not welded and the open portion 13 remains open, the EO gas is quickly discharged without remaining in the culture bag 10.
- evacuation air is supplied to perform air replacement. Then, such evacuation and air replacement are further performed twice, and aeration is performed three times in total. Then, open the door of the sterilizer, take out the product, and check the discoloration of the indicator. If the discoloration of the indicator is confirmed, the sterilization process is completed.
- FIG. 7 is an example of a diagram showing a state in which the culture bag 10 containing the culture medium 20 is sealed.
- the sterilization bag 30 containing the culture bag 10 is kept sealed after the sterilization gas in the culture bag 10 is removed.
- the open portion 13 of the culture bag 10 is closed from the outside of the sterilization bag 30 with the welding sealer 2, and the culture bag 10 containing the sterilized medium 20 is sealed.
- the culture bag 10 will be in the state by which two sheet-like raw materials which comprise the culture bag 10 weld each other in the edge part 12B shown in FIG. 1, and the culture bag 10 was sealed.
- the treatment for removing the sterilization gas in the culture bag 10 includes, for example, the sterilization bag 30 containing the culture bag 10 in a non-sterile gas atmosphere after the sterilization process and before the start of the sealing process of the culture bag 10.
- a method of placing the sterilization bag 30 in a state where the culture bag 10 is housed in a tank capable of being evacuated for vacuuming and placing the sterilization bag 30 in a low-pressure atmosphere. is there.
- FIG. 8 is an example of a diagram showing a sterilization bag 30 containing the culture bag 10 after the open portion 13 is closed with the welding sealer 2.
- the inside of the culture bag 10 is closed over the entire circumference by the edge portions 12A and 12B. Accordingly, the culture medium 20 in the culture bag 10 is isolated from microorganisms and other foreign matters outside the culture bag 10.
- FIG. 9 is an example of a cross-sectional view of the sterilization bag 30 and the culture bag 10 taken along the line indicated by the reference numeral BB in FIG.
- the culture bag 10 has a liquid impermeability in which each of the two sheet-like materials constituting the culture bag 10 does not leak a sample and an oxygen permeability that transmits oxygen necessary for the growth of microorganisms, and While either one of the materials can be fused to the other material, the film 33 constituting the sterilization bag 30 is formed so as not to be fused.
- the material constituting the outer surface of the film 33 of the sterilization bag 30 is “material 1 (outside)”, the material constituting the inner surface of the film 33 is “material 1 (inside)”, and the sheet 31 of the sterilization bag 30 is configured.
- “Material 2” the material constituting the outer surface of the film 14 of the culture bag 10 “Material 3 (outside)”, the material constituting the inner surface of the film 14 “Material 3 (inside)”, the culture bag
- the melting point of each material is as follows: By satisfying the relationship, the temperature of the welding sealer 2 when closing the open portion 13 is appropriately adjusted, and the open portion 13 of the culture bag 10 is sterilized without fusing the culture bag 10 to the sterilization bag 30.
- the microorganism incubator 1 is completed through the above series of manufacturing steps. Since the microorganism culture device 1 according to the present embodiment is completed through the manufacturing process of the above example, for example, the culture bag is sterilized with the sterilization bag 30 omitted, and then the open portion of the culture bag is closed. Compared to the case, the possibility that fungi and other foreign substances enter the culture bag can be greatly suppressed.
- the microorganism incubator 1 that has been completed through the series of manufacturing steps described above is shipped, for example, while the culture bag 10 is stored in the sterilization bag 30, so that the surface of the culture bag 10 in the distribution process can be obtained. It is possible to suppress the adhesion of microorganisms and other foreign matters to the skin. If the adhesion of foreign matter to the surface of the culture bag 10 is suppressed, the possibility of foreign matter entering the culture bag 10 when the injection port 11 is opened decreases.
- the above production method is not limited to the mode in which the culture medium 20 is put into the culture bag 10 from the open portion 13 provided on the opposite side of the inlet 11 when the culture medium 20 is put into the culture bag 10.
- the culture medium 20 may be put into the culture bag 10 from an open portion provided in the vicinity of the injection port 11 or in other portions.
- the above manufacturing method is not limited to an embodiment in which the culture bag 10 is placed in the sterilization bag 30 from the injection port 11 side.
- the culture bag 10 is inserted from the open portion 13 side into the sterilization bag 30. You may put it in.
- the culture medium 20 may be stored in the culture bag 10 by the following method, for example.
- two sheets of sheet-shaped material constituting the culture bag 10 are stacked, the medium 20 is sandwiched between them, and the edges of the sheet-shaped material are heated to weld the edge portion 12A. May be stored in the culture bag 10.
- the culture bag 10 is not limited to the one in which the edges of the sheet-like material are heated and welded to each other, and may be formed by bonding with an adhesive tape or the like, for example.
- the sterilization bag 30 is realizable by using the material shown in the following Table 1, for example.
- the sterilization bag 30 may be provided with gas permeability to the sheet 31 by using, for example, the material shown in Table 1 above, and the film 33 may be provided with heat meltability that can be fused to the sheet 31. Is possible.
- the culture bag 10 is realizable by using the material shown in the following Table 2, for example.
- “Material 3 (outside)” corresponds to the outside (the side not touching the film 15) portion of the film 14 shown in FIG. 9, and “Material 3 (inside)” is the film shown in FIG. 14 corresponds to the inner part of the film 15 (the side touching the film 15), and the “material 4 (inner side)” corresponds to the inner part of the film 15 shown in FIG. "Outside” "corresponds to the outside (the side not touching the film 14) portion of the film 15 shown in FIG.
- the material which comprises the culture bag 10 is not limited to the aspect shown by the said Table 2, For example, the outside of PET / PE and the inside are OPP / PE, etc. It is also possible to further modify the combination.
- the culture bag 10 is made of a material having an oxygen permeability and a melting point as shown in Tables 3, 4 and 5 above, for example, in combination with a pattern as shown in Table 2 above, thereby leaking the sample. Sterilization while providing film 14 and 15 with a liquid impermeability and oxygen permeability that allows oxygen necessary for microbial growth, and either film can be fused to the other film. It is possible to give the films 14 and 15 heat melting property having a melting point lower than that of the film 33 constituting the bag 30.
- the culture medium 20 accommodated in the culture bag 10 is preferably a laminate suitable for the growth of microorganisms.
- a sheet-shaped culture medium including a porous matrix layer and a water-soluble polymer compound layer is suitable.
- the culture medium 20 for example, there is one including a substrate, a water-soluble polymer compound layer, and a porous matrix layer.
- the water-soluble polymer layer can contain nutrients suitable for microorganisms intended for culture.
- the water-soluble polymer layer further includes a pH adjuster, a selective agent for suppressing the growth of microorganisms other than the intended purpose, a color former, a dye, and a surfactant for making the growth of microorganisms easier to see or confirming the growth of specific microorganisms.
- a pH adjuster a selective agent for suppressing the growth of microorganisms other than the intended purpose
- a color former e.g., a dye, and a surfactant for making the growth of microorganisms easier to see or confirming the growth of specific microorganisms.
- At least one selected from inorganic salts and the like can be included.
- Examples of the method for producing the medium 20 include a water-soluble polymer compound, a nutritional component suitable for microorganisms intended for culture, a pH adjuster, a selective agent that suppresses the growth of microorganisms other than the target, and the growth of microorganisms is easy to see.
- a method in which an aqueous solution containing a color former, a dye, a surfactant, an inorganic salt, etc. for confirming the growth of a specific microorganism is applied on a substrate and dried to form a film.
- An aqueous solution of a water-soluble polymer compound that is separately contained may be sequentially applied and overlaid.
- the culture medium 20 includes a porous matrix layer and at least one water-soluble polymer compound layer, and at least one of the water-soluble polymer compound layers includes a nutrient component.
- a specific method for producing the culture medium 20 is, for example, a method in which an aqueous solution of a water-soluble polymer compound containing a nutrient component, a salt component, and a chromogenic enzyme substrate is applied onto a film (substrate) such as polyester to form a dry film.
- a film substrate
- a chromogenic enzyme substrate is applied onto a film (substrate) such as polyester to form a dry film.
- an aqueous solution of a water-soluble polymer is layered on a film (substrate) of polyester or the like, formed into a dry film, and then an aqueous solution of a water-soluble polymer containing nutritional components and the like is formed on the film. Multilayer and dry film. Further, an aqueous solution of a water-soluble polymer containing a color former (this solution may further contain a nutrient component and a pH adjuster) is layered on this film to form a dry film. And the method of laminating
- the porous matrix When the porous matrix is overlaid, the porous matrix may be overlaid before the top layer is dried and then dried. You may apply
- the dry film-like medium layer produced in this manner is peeled off from a film (substrate) such as polyester, or is not peeled off, and is commensurate with the amount of sample liquid to be cultured.
- a sheet-like culture medium 20 prepared by cutting to a predetermined size is prepared, and the above-described series of manufacturing steps is performed.
- the substrate of the medium 20 is not particularly limited as long as the tensile strength at the time of heating and drying is sufficient, but generally, for example, a polyester film having a thickness of 20 to 50 ⁇ m is used.
- the water-soluble polymer compound forming the water-soluble polymer compound layer of the medium 20 may be any material that dissolves when a sample is added and exhibits a viscosity of 10 cps or more and does not inhibit the growth of microorganisms.
- polyvinyl alcohol modified polyvinyl alcohol (in the polymerization of vinyl acetate, unsaturated dicarboxylic acid (for example, maleic acid, fumaric acid, glutaconic acid, allylmalonic acid, their anhydrides, or monoalkyl esters thereof) Polymerized (for example, see Japanese Examined Patent Publication No.
- polyvinyl alcohol reacted with cyclic acid anhydride polyvinyl alcohol reacted with cyclic acid anhydride; cellulose derivative (for example, carboxymethyl cellulose (CMC), hydroxyalkyl cellulose (for example, hydroxy Starch and derivatives thereof (eg soluble starch, carboxymethylated starch); polysaccharides other than cellulose derivatives, starch and derivatives thereof (eg hyaluronic acid, guar gum, gum arabic); acrylic acid and derivatives thereof ( Eg to polyacrylic acid, polyacrylates, acrylic acid (salt) - vinyl alcohol copolymers); polyethers (e.g., polyethylene glycol, polypropylene glycol); proteins, protein derivatives (e.g., collagen) and the like.
- CMC carboxymethyl cellulose
- hydroxyalkyl cellulose for example, hydroxy Starch and derivatives thereof (eg soluble starch, carboxymethylated starch)
- the water-soluble polymer compound has a viscosity of 4 mass% aqueous solution measured at 20 ° C. using an Ostwald viscometer, preferably 10 cps or more, more preferably 15 to 80 cps.
- a water-soluble polymer compound By using such a water-soluble polymer compound, the microorganism does not enter the inside of the medium, and the microorganism that has started to split does not move on the surface of the medium. This is preferable because counting becomes easy.
- these water-soluble polymer compounds those selected from the group consisting of cellulose derivatives and polyvinyl alcohol are preferable, and polyvinyl alcohol having a saponification degree of 75 to 95% and a molecular weight of 25,000 to 250,000 is particularly preferable.
- the suitable content of the water-soluble polymer compound in the culture medium 20 of the microorganism culture device 1 according to this embodiment is suitably 40 to 300 g / m 2 , and preferably 70 to 150 g / m 2 .
- Synthetic fibers for example, nylon, polyacrylonitrile, polyester (especially hydrophilized), polyolefins (especially hydrophilized polyurethane)
- semi-synthetic fibers as materials for forming the porous matrix layer of the culture medium 20
- rayon, etc. Natural fibers (for example, wool (animal hair), silk, cotton, cellulose, pulp, etc.), woven fabrics, fiber webs, non-woven fabrics, etc. formed of inorganic fibers (for example, glass fibers), etc.
- examples thereof include porous films, sponges, and porous ceramics made of fiber constituent materials.
- the porous matrix does not necessarily need to be hydrophilic.
- porous matrix or a porous matrix subjected to hydrophilic treatment
- the water absorption speed is increased and the work efficiency is increased.
- a water-soluble substance to the surface of the hydrophilic or hydrophilic-treated porous matrix because the dispersibility of microorganisms is improved.
- the porous matrices fiber webs and nonwoven fabrics are preferred.
- Nylon, cotton, cellulose, and rayon are preferable as the fibers constituting the fiber web and the nonwoven fabric, and a fiber web or nonwoven fabric made of nylon is particularly preferable.
- Nylon nonwoven fabric is preferably a nonwoven fabric produced by a melt blown manufacturing method.
- porous matrix layer is a nylon melt blown nonwoven fabric having a basis weight of 40 to 100 g / m 2 and an air permeability of 7 to 24 cm / sec.
- Specific examples of the porous matrix layer include those disclosed in WO 01/44437.
- the nutrient component used in the culture medium 20, that is, the culture medium component may be any material as long as it is suitable for the growth of the microorganism targeted for detection.
- a medium component obtained by removing agar from a general-purpose liquid medium or agar medium can be used.
- a selective agent that suppresses the growth of microorganisms other than the detection purpose, an indicator for making the generated colonies easy to see, a coloring agent, etc. may be added to the sheet medium, and a coloring or fluorescent enzyme may be used to detect specific microorganisms.
- Substrate may be added.
- pigments, surfactants, inorganic salts, and the like can be added.
- medium components for microorganisms include, for example, yeast extract / peptone / glucose mixture, meat extract / pepton mixture, peptone / soybean peptone / glucose mixture, phosphate phosphate, sodium chloride , Carbonates, magnesium salts, and other salts are added to test for Escherichia coli and coliforms.
- Sodium desoxycholate / pepton / ammonium iron citrate / sodium chloride / dipotassium phosphate / lactose / neutral red, peptone ⁇ Lactose, dipotassium phosphate, eosin Y, methylene blue, etc. are used for staphylococcal testing.
- Yellow extract such as tellurite egg yolk mixture
- Yellow extract such as tellurite egg yolk mixture
- Yeast extract, peptone, sucrose, sodium thiosulfate, sodium citrate, sodium cholate, ferric citrate, sodium chloride, beef bile, bromthymol blue -Thymol blue mixture, etc. for enterococci include cattle brain extract, heart extract, peptone, glucose, dipotassium phosphate, sodium nitride, bromthymol blue, 2,3,5-triphenyltetrazolium chloride mixture, etc.
- peptone / glucose mixture for fungi, peptone / glucose mixture, yeast extract / glucose mixture, potato extract / glucose mixture, etc., malt extract / pepton / glucose mixture, etc. for salmonella, peptone, meat extract, glycerin, sodium thiosulfate, L -Lysine, ammonium ammonium citrate, deoxy Sodium oxalate, sodium lauryl sulfate mixture, etc. for aqueous bacteria include peptone, yeast extract, casamino acid, glucose, phosphate, soluble starch, magnesium salt mixture, meat extract or fish meat extract, tryptone, glucose Etc. are used.
- pH adjusters examples include phosphates and carbonates (monosodium phosphate, disodium phosphate, monopotassium phosphate, dipotassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate, calcium carbonate, etc.) Is mentioned. Such a pH adjuster is used to maintain a pH suitable for the growth of microorganisms.
- antibiotics such as antibiotics and synthetic antibacterial agents, pigments, surfactants, inorganic salts and the like are used.
- antibiotics include methicillin, ceftamethazole, cefixime, ceftazidime, cefsulosin, bacitracin, polymyxin B, rifampicin, novobiocin, colistin, lincomycin, chloramphenicol, tetracycline, streptomycin, etc.
- examples thereof include nalidixic acid and olaquindox.
- Examples of the pigment include bacteriostatic or bactericidal crystal violet, brilliant green, malachite green, and methylene blue.
- Examples of the surfactant include Tergitol 7, dodecyl sulfate, lauryl sulfate and the like.
- examples of inorganic salts include selenite, tellurite, sulfite, sodium nitride, lithium chloride, oxalate, and high-concentration sodium chloride. Besides these, taurocholate, glycine, bile powder, bile salt, deoxycholate, and the like can be used.
- a color or fluorescent enzyme substrate such as tetrazolim salt or esterase or phosphatase
- the growth of microorganisms can be easily observed as a color or fluorescent spot.
- a specific microorganism can be detected by adding a coloring or fluorescent enzyme substrate of an enzyme possessed by the specific microorganism.
- a coloring or fluorescent enzyme substrate of an enzyme possessed by the specific microorganism For example, 5-bromo-4-chloro-3-indolyl phosphate, 2,3,5-triphenyltetrazolium chloride, 6-chloro-3-indoxyl- ⁇ -D-glucuronic acid and the like.
- the nutrient component originally contained in the dialysate is not included in order to prevent the microorganisms in the dialysate or reverse osmosis water from dying due to overnutrition. It is sufficient if it has a supplementary nutrient component required for microbes in the reverse osmosis water, such as 0.1 to 3.0 g peptone and 0.1 to 3.0 g per square meter.
- Yeast extract 0.1-0.5 g casein degradation product, 0-0.5 g glucose, 0.075-0.3 g sodium pyruvate, and 0-0.3 g dipotassium phosphate And 0.1-0.5 g soluble starch, 0.01-0.05 g magnesium sulfate, 0.015-0.1 g tetrazolium salt or 0.15-1.0 g indoxyl derivative. thing It can be.
- the medium components are 0.1 to 3.0 g of peptone, 0.1 to 3.0 g of yeast extract, and 0.1 to 0.5 g.
- Casein degradation product 0-0.5 g glucose, 0.075-0.3 g sodium pyruvate, 0-0.3 g dipotassium phosphate, 0.1-0.5 g soluble starch 0.01 to 0.05 g of magnesium sulfate, and 0 to 0.3 g of dipotassium phosphate is a component that serves as a pH adjuster. If the above range is exceeded, the detection speed and sensitivity may be reduced or undetectable due to a decrease in the growth rate of bacteria, a decrease in the number of viable bacteria, and the like.
- the chromogenic substrate (component for making the grown bacteria easy to see) is 0.015-0.1 g of tetrazolium salt or 0.15-1.0 g of indoxyl derivative, and the above lower limit If it exceeds, it will be difficult to detect visually. In addition, when the above upper limit is exceeded, the growth of bacteria is inhibited and the detection sensitivity is lowered. Alternatively, the background is increased due to excessive coloration of the substrate. As a result, it becomes difficult to distinguish the number of bacteria.
- the nutrient component originally contained in the dialysate is not included in order to prevent the microorganisms in the dialysate or reverse osmosis water from dying due to overnutrition.
- a certain supplementary nutritional component necessary for the microbes in the reverse osmosis water such as 0.38 to 9.0 g of meat extract or fish extract per square meter, and 0.63 ⁇ 15.0 g tryptone, 0 ⁇ 3.0 g glucose, 0 ⁇ 0.3 g dipotassium phosphate, 0.015 ⁇ 0.1 g tetrazolium salt or 0.15 ⁇ 1.0 g indoxyl Derivatives can be mentioned.
- the medium components are 0.38 to 9.0 g of meat extract or fish extract, 0.63 to 15.0 g of tryptone, and 0 to 3. It is 0 g of glucose, and 0 to 0.3 g of dipotassium phosphate is a component that serves as a pH adjuster. If the above range is exceeded, the detection speed and sensitivity may be reduced or undetectable due to a decrease in the growth rate of bacteria, a decrease in the number of viable bacteria, and the like.
- the chromogenic substrate (component for making the grown bacteria easy to see) is 0.015 to 0.1 g of tetrazolium salt or 0.15 to 1.0 g of indoxyl derivative. If it exceeds, it will be difficult to detect visually. In addition, when the above upper limit is exceeded, the growth of bacteria is inhibited and the detection sensitivity is lowered. Alternatively, the background is increased due to excessive coloration of the substrate. As a result, it becomes difficult to distinguish the number of bacteria.
- the nutrient component originally contained in the dialysate is not included in order to prevent the microorganisms in the dialysate or reverse osmosis water from dying due to overnutrition. What is necessary is that it has a predetermined supplementary nutrient component necessary for microbes in the reverse osmosis water, such as 2.5 to 40.0 g of malt extract and 0 to 40.0 g per square meter.
- Glucose 0.25-4.0 g peptone, 0-0.3 g monopotassium phosphate, 0-0.1 g chloramphenicol, 0.015-0.1 g tetrazolium salt or 0 15 to 1.0 g of indoxyl derivatives.
- the medium components are 2.5 to 40.0 g of malt extract, 0 to 40.0 g of glucose, and 0.25 to 4.0 g of peptone.
- From 0 to 0.3 g of monopotassium phosphate is a component that serves as a pH adjuster.
- 0 to 0.1 g of chloramphenicol is a component that serves as a selective agent. If the above range is exceeded, the detection speed and sensitivity may be reduced or undetectable due to a decrease in the growth rate of bacteria, a decrease in the number of viable bacteria, and the like.
- the chromogenic substrate (component for making the grown bacteria easy to see) is 0.015-0.1 g of tetrazolium salt or 0.15-1.0 g of indoxyl derivative, and the above lower limit If it exceeds, it will be difficult to detect visually. In addition, when the above upper limit is exceeded, the growth of bacteria is inhibited and the detection sensitivity is lowered. Alternatively, the background is increased due to excessive coloration of the substrate. As a result, it becomes difficult to distinguish the number of bacteria.
- the shape of the culture medium 20 is, for example, a square such as a square or a rectangle, the production is easy.
- the area and thickness of the medium 20 are appropriately determined depending on the amount of the sample solution to be inspected, the water-soluble polymer compound used in the sheet-shaped medium, the porous matrix, and their amounts.
- the medium 20 desirably satisfies the relationship between the mass of the water-soluble polymer compound represented by the following formula (1) and the amount of the sample liquid as the specimen, and contains the water-soluble polymer compound in the sheet-shaped medium as described above.
- the size is preferably such that the amount is 40 to 300 g / m 2 .
- a water-soluble polymer compound layer, a water-soluble polymer compound layer containing a nutrient component, a water-soluble polymer compound layer, and a porous matrix layer are arranged in this order on a substrate (for example, a polyester film).
- a laminated layer can be mentioned.
- a water-soluble polymer compound layer, a water-soluble polymer compound layer containing a nutrient component, a water-soluble polymer compound layer containing a color former, and a porous matrix layer on a substrate (for example, a polyester film) are stacked in this order.
- the culture medium 20 of the microorganism culture device 1 for example, the microorganism culture device disclosed in International Publication No. 97/24432 pamphlet can be used.
- the conditions for preserving the microorganism incubator 1 before use containing the medium 20 can be appropriately selected depending on the types and stability of nutrient components and color formers. It is also possible to select.
- FIG. 10 is an example of a diagram illustrating a state in which the culture bag 10 is taken out from the sterilization bag 30.
- FIG. 11 is an example of a diagram showing a state in which a sample is put into the culture bag 10.
- the culture bag 10 is taken out from the sterilization bag 30 as shown in FIG. 10.
- the inlet 11 of the culture bag 10 is opened, the sample is placed in the culture bag 10, and the inlet 11 is closed again.
- the mass (g) of the water-soluble polymer compound in the medium 20 and the volume (mL) of the sample solution have the relationship of the above formula (1).
- the culture bag 10 is placed under conditions such as temperature and time suitable for the microorganism to be cultured, and cultivation of the microorganism is started.
- the microorganism culture device 1 according to the present embodiment is suitable for aerobic bacteria culture, anaerobic bacteria can be cultured by forming the culture bag 10 from an oxygen-impermeable material. It is.
- the sample does not leak at the time of addition of the sample, is liquid impervious, and has oxygen impermeability to grow anaerobic bacteria. It is desirable. Further, it is preferably transparent or translucent in order to observe the growth of microorganisms. As a material satisfying such requirements, for example, substantially non-porous, it is preferable oxygen permeability is 0 ⁇ 20mL / m 2/24 hours.
- microorganisms cultured by the above method By examining the presence or amount of microorganisms cultured by the above method, various microorganisms can be inspected.
- the culture bag 10 can also change the position of the inlet 11 suitably as shown, for example in FIGS. 12-15.
- the injection port 11 is not limited to a simple screw-type cap as shown in FIG. 16, for example, and any sample can be used as long as it can inject a sample and open / close the injection port 11. Also good.
- caps other than the cap applicable as the inlet 11 include, for example, spouts, screw caps, push-in caps, crowns, rubber stoppers, urethane stoppers, silicone stoppers, cork stoppers, molton stoppers, cotton stoppers, paper stoppers, three-way cocks. , Fasteners, chucks, and zippers. That is, for example, a one-touch type cap as shown in FIG. 17, a rubber-type lid for inserting a sample with an injection needle or the like as shown in FIG.
- Example 1 of medium 15 g of polyvinyl alcohol having a saponification degree of 89% and a polymerization degree of 1700 was added to 0.125 L of water, dissolved by heating, and then applied onto a 20 ⁇ m thick 0.5 ⁇ 0.5 m polyester film, and dried at 120 ° C. for 5 minutes to form a film.
- Nylon melt blown nonwoven fabrics having a basis weight of 65 g / m 2 and an air permeability of 110 L / m 2 sec were laminated and dried at 100 ° C. for 30 seconds.
- the sheet-like culture medium 20 is cut into 120 ⁇ 150 mm and placed in the culture bag 10, the culture bag 10 containing the culture medium 20 is placed in the sterilization bag 30, the open portion 32 is closed, sterilized with ethylene oxide gas, and opened. The portion 13 was closed to produce the microorganism incubator 1.
- the culture bag 20 used here is made of two nylon / LLDPE (Linear Low Density Polyethylene) films (size: 150 ⁇ 220 mm), and the edge portion 12A is sandwiched between the spouts serving as the injection ports 11. Welded.
- the sterilization bag 30 used here is a sterilization rollback (size: 200 ⁇ 350) of Medicom (Medicom is a registered trademark).
- the culture medium 20 which concerns on a present Example has low glucose concentration and sodium carbonate density
- a nutrient component such as a yeast extract. If nutrients become excessive, the growth of bacteria may be inhibited due to reasons such as an increase in osmotic pressure against bacteria, but the medium 20 according to the present embodiment lowers the glucose concentration and makes the sodium carbonate concentration zero. Therefore, even if a nutrient component is added, it is difficult to inhibit growth.
- Nylon melt blown nonwoven fabrics having a basis weight of 65 g / m 2 and an air permeability of 110 L / m 2 sec were laminated and dried at 100 ° C. for 30 seconds.
- the sheet-like culture medium 20 is cut into 120 ⁇ 150 mm and placed in the culture bag 10, the culture bag 10 containing the culture medium 20 is placed in the sterilization bag 30, the open portion 32 is closed, sterilized with ethylene oxide gas, and opened. The portion 13 was closed to produce the microorganism incubator 1.
- the culture bag 20 used here is made of two nylon / LLDPE (Linear Low Density Polyethylene) films (size: 150 ⁇ 220 mm), and the edge portion 12A is sandwiched between the spouts serving as the injection ports 11. Welded.
- the sterilization bag 30 used here is a sterilization rollback (size: 200 ⁇ 350) of Medicom (Medicom is a registered trademark).
- the sheet-like culture medium 20 is cut into 120 ⁇ 150 mm and placed in the culture bag 10, the culture bag 10 containing the culture medium 20 is placed in the sterilization bag 30, the open portion 32 is closed, sterilized with ethylene oxide gas, and opened. The portion 13 was closed to produce the microorganism incubator 1.
- the culture bag 20 used here is made of two nylon / LLDPE (Linear Low Density Polyethylene) films (size: 150 ⁇ 220 mm), and the edge portion 12A is sandwiched between the spouts serving as the injection ports 11. Welded.
- the sterilization bag 30 used here is a sterilization rollback (size: 200 ⁇ 350) manufactured by Medicom (Medicom is a registered trademark).
- ⁇ Comparison with MF method and simplified MF method> In relation to the examination of the dialysate, the method using the microorganism culture device 1 according to the above embodiment was compared with the conventional MF method. In this comparison, a bacterial solution was diluted with a dialysis solution to prepare about 4 CFU / mL. 25 mL of this bacterial solution was added to each of the microorganism incubator 1 according to the above embodiment and the filter of the MF method. In the MF method, the bacterial solution was suction filtered and the filter was transferred onto an R2A agar medium. The number of bacteria was measured after 3 days and 5 days after culturing at 25 ° C. (only Methylobacterium extorquens was measured after 7 days). The results are shown below.
- the method using the microorganism culture device 1 according to the above embodiment was compared with the conventional simple MF method.
- a bacterial solution was diluted with a dialysis solution to prepare about 0.4 CFU / mL. 25 mL of this bacterial solution was added to each of the microorganism incubator 1 according to the above embodiment and a conventional simple MF filter.
- the simple MF method the bacterial solution was suction filtered, and 2 mL of mTGE medium was added and suction filtered. The number of bacteria was measured after 3 days and 5 days after culturing at 25 ° C. (only Methylobacterium extorquens was measured after 7 days). The results are shown below.
- Tables 6 and 7 above are for the case where the medium according to Example 1 is used, but the data when the medium according to Example 2 is used are as follows.
- the data when the medium according to Example 3 is used is as shown in Table 9 below.
- Tables 6 and 7 the numbers of bacteria after 3 days and 5 days were shown, but in Tables 8 and 9 below, the numbers of bacteria after 3 days and 6 days were shown.
- the number of bacteria was confirmed to be larger when the microorganism culture device 1 according to the above embodiment was used than when the conventional MF method was used. It can be seen that Moreover, about the culture medium which concerns on Example 2, it turns out that a substantially equivalent bacterial count is confirmed by the case where the microorganisms culture device 1 which concerns on the said embodiment is used, and the case where the conventional MF method is used. .
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Abstract
Description
図1は、実施形態に係る微生物培養器の外観図の一例である。微生物培養器1は、図1に示すように、袋状の培養袋10にシート状の培地20を入れた微生物培養器であり、滅菌袋30に収容されている。なお、微生物培養器1は、培養袋10を収容する滅菌袋30が備わっているものに限定されるものでなく、滅菌袋30は省略されていてもよい。微生物培養器1は、細菌を検査する際にあると便利なものを適宜含めた微生物検査キットの全部または一部を構成するものであってもよい。培養袋10には、試料を容器内に注入するための注入口11が取り付けられている。培養袋10は、2枚重ねにしたシート状の素材(例えば、Polymer等の熱融解性材料)の縁同士を溶着したものであり、図1に示すように、縁の部分12A,12Bが溶着されている。滅菌袋30についても、培養袋10と同様、2枚重ねにしたシート状の素材の縁同士を溶着したものであり、図1に示すように、縁の部分31A,31Bが溶着されている。
微生物培養器1は、例えば、以下のような製造工程により製造可能である。
「素材3,4(内側)」≪「素材1(内側)」≪「素材1(外側),素材2,素材3,4(外側)」
なお、培養袋10に収容する培地20は、微生物の生育に適する積層物であることが望ましく、例えば、多孔質マトリックス層と水溶性高分子化合物層とを含むシート状の培地が好適である。培地20のさらなる具体的構造としては、例えば、基板、水溶性高分子化合物層及び多孔質マトリックス層を含むものがある。
式(1):0.08<水溶性高分子化合物の重量(g)/試料液量(mL)<0.5
図10は、滅菌袋30から培養袋10を取り出す様子を示した図の一例である。また、図11は、培養袋10内に試料を入れる様子を示した図の一例である。微生物培養器1を使用する際は、図10に示すように、滅菌袋30から培養袋10を取り出す。このとき、滅菌袋30に保護されていた培養袋10の表面に異物類がなるべく付着しないように配慮することが好ましい。次に、図11に示すように、培養袋10の注入口11を開いて試料を培養袋10内に入れ、注入口11を再び閉じる。培養にあたっては、培地20における水溶性高分子化合物の質量(g)と試料液の容量(mL)とが上記式(1)の関係にあることが望ましい。注入口11を閉じて培養袋10内を密閉した後は、培養袋10を、培養しようとする微生物に適した温度及び時間などの条件下に置き、微生物の培養を開始する。なお、本実施形態に係る微生物培養器1は、好気性菌の培養に適しているが、培養袋10を酸素不透過性の素材で形成することにより、嫌気性菌の培養を行うことも可能である。嫌気性菌の検出用として用いる場合の培養袋10の材質としては試料の添加時に該試料が漏れず、液体不浸透性であって、一方嫌気性細菌が生育する酸素不透過性を有していることが望ましい。また、微生物の生育を観察するために透明または半透明であることが好ましい。このような要求を満たす材質としては、例えば、実質的に無孔性であり、酸素透過率が0~20mL/m2/24時間であることが好ましい。
なお、培養袋10は、例えば、図12から図15に示すように、注入口11の位置を適宜変更することも可能である。
水0.125Lに鹸化度89%重合度1700のポリビニルアルコール15gを加え加熱溶解後、20μm厚0.5×0.5mのポリエステルフィルム上に塗布し、120℃、5分間乾燥しフィルム化した。このフィルム上に、鹸化度89%、重合度1700のポリビニルアルコール8g、ペプトン0.0625g、酵母エキス0.0625g、カザミノ酸0.0625g、グルコース0.015625g、リン酸二カリウム0.0375g、溶性デンプン0.0625g、硫酸マグネシウム0.00625gを水0.125Lに溶解して、110℃、7分間乾燥した。このフィルム上に、ポリビニルアルコール2.5g、5-ブロモ-4-クロロ-3-インドリルフォスフェート0.075g、2,3,5-トリフェニルテトラゾリウムクロライド0.0075gを水0.05Lに溶解して重層し、目付65g/m2、通気度110L/m2secのナイロンメルトブロー不織布を張り合わせ100℃30秒間乾燥した。
水0.125Lに鹸化度89%重合度1700のポリビニルアルコール15gを加え加熱溶解後、20μm厚0.5×0.5mのポリエステルフィルム上に塗布し、120℃、5分間乾燥しフィルム化した。このフィルム上に、鹸化度89%、重合度1700のポリビニルアルコール8g、肉エキス0.375g、トリプトン0.625g、グルコース0.125gを水0.125Lに溶解して、110℃、7分間乾燥した。このフィルム上に、ポリビニルアルコール2.5g、5-ブロモ-4-クロロ-3-インドリルフォスフェート0.075g、2,3,5-トリフェニルテトラゾリウムクロライド0.0075gを水0.05Lに溶解して重層し、目付65g/m2、通気度110L/m2secのナイロンメルトブロー不織布を張り合わせ100℃30秒間乾燥した。
水0.125Lに鹸化度89%重合度1700のポリビニルアルコール15gを加え加熱溶解後、20μm厚0.5×0.5mのポリエステルフィルム上に塗布し、120℃、5分間乾燥しフィルム化した。このフィルム上に、鹸化度89%、重合度1700のポリビニルアルコール8g、麦芽エキス2.5g、グルコース2.5g、ペプトン0.125g、リン酸一カリウム0.0375g、クロラムフェニコール0.0125gを水0.125Lに溶解して、110℃、7分間乾燥した。このフィルム上に、ポリビニルアルコール2.5g、5-ブロモ-4-クロロ-3-インドリルフォスフェート0.075g、2,3,5-トリフェニルテトラゾリウムクロライド0.0075gを水0.05Lに溶解して重層し、目付65g/m2、通気度110L/m2secのナイロンメルトブロー不織布を張り合わせ100℃30秒間乾燥した。
透析液の検査に関連し、上記実施形態に係る微生物培養器1を用いた方法と、従来からあるMF法との比較を行った。本比較においては、菌液を透析液で希釈して約4CFU/mLとしたものを用意した。この菌液を、上記実施形態に係る微生物培養器1とMF法のフィルタとにそれぞれ25mL添加した。MF法では、菌液を吸引ろ過後、フィルタをR2A寒天培地上に移した。それぞれ25℃で培養して3日後、5日後の菌数を測定した(Methylobacterium extorquensのみ7日後も測定した)。その結果を以下に示す。
Claims (17)
- シート状の培地を内部に入れて封止した注入口付の培養袋を含む、微生物培養器。
- 培養袋の注入口は、スパウト、スクリューキャップ、押込キャップ、王冠、ゴム栓、ウレタン栓、シリコン栓、コルク栓、モルトン栓、綿栓、紙栓、3方コック、ファスナー、チャック、ジッパーの何れか1種類以上であることを特徴とする、請求項1に記載の微生物培養器。
- シート状の培地を内部に入れて封止した注入口付の培養袋と、
培養袋を内部に入れて封止した滅菌袋と、を備えることを特徴とする、請求項1または2に記載の微生物培養器。 - 微生物培養器は、好気性菌の検出用であり、
培養袋は、酸素透過性の素材で形成されていることを特徴とする、請求項1から3の何れか一項に記載の微生物培養器。 - 微生物培養器は、嫌気性菌の検出用であり、
培養袋は、酸素不透過性の素材で形成されていることを特徴とする、請求項1から3の何れか一項に記載の微生物培養器。 - 微生物培養器は、透析液中または逆浸透水中に含まれる微生物の検出用であり、
培養袋内の培地は、透析液中または逆浸透水中に含まれている栄養成分を補う所定の補助栄養成分を有することを特徴とする、請求項1から4の何れか一項に記載の微生物培養器。 - 培養袋内の培地が有する所定の補助栄養成分とは、一平方メートルあたり0.1~3.0gのペプトンと、0.1~3.0gの酵母エキスと、0.1~0.5gのカゼイン分解物と、0~0.5gのグルコースと、0.075~0.3gのピルビン酸ナトリウムと、0~0.3gのリン酸二カリウムと、0.1~0.5gの溶性デンプンと、0.01~0.05gの硫酸マグネシウムと、0.015~0.1gのテトラゾリウム塩または0.15~1.0gのインドキシル誘導体であることを特徴とする、請求項6に記載の微生物培養器。
- 培養袋内の培地が有する所定の補助栄養成分とは、一平方メートルあたり0.38~9.0gの肉エキスまたは魚肉エキスと、0.63~15.0gのトリプトンと、0~3.0gのグルコースと、0~0.3gのリン酸二カリウムと、0.015~0.1gのテトラゾリウム塩または0.15~1.0gのインドキシル誘導体であることを特徴とする請求項6に記載の微生物培養器。
- 培養袋内の培地が有する所定の補助栄養成分とは、一平方メートルあたり2.5~40.0gの麦芽エキスと、0~40.0gのグルコースと、0.25~4.0gのペプトンと、0~0.3gのリン酸一カリウムと、0~0.1gのクロラムフェニコールと、0.015~0.1gのテトラゾリウム塩または0.15~1.0gのインドキシル誘導体であることを特徴とする請求項6に記載の微生物培養器。
- 請求項1から9の何れか一項に記載の微生物培養器を含む微生物検査キット。
- シート状の培地を内部に入れて封止した注入口付の培養袋と、培養袋を内部に入れて封止した滅菌袋とを備える微生物培養器の、滅菌袋に封止されている培養袋を滅菌袋から取り出し、
培養袋の注入口を開いて注入口から培養袋内へ透析液を入れ、
注入口を密閉してから培養袋を微生物培養の条件下に置いて微生物を培養し、
培養した微生物の存在又は量を検査する、
透析液の検査方法。 - シート状の培地を内部に入れて封止した注入口付の培養袋と、培養袋を内部に入れて封止した滅菌袋とを備える微生物培養器の、滅菌袋に封止されている培養袋を滅菌袋から取り出し、
培養袋の注入口を開いて注入口から培養袋内へ試料液を入れ、
注入口を密閉してから培養袋を微生物培養の条件下に置く、
微生物の培養方法。 - 請求項12に記載の微生物の培養方法によって培養した微生物の存在又は量を検査する微生物の検査方法。
- シート状の培地を内部に入れて封止した注入口付の培養袋と、培養袋を内部に入れて封止した滅菌袋とを備える微生物培養器の製造方法であって、
培地を培養袋の開封部分から内部に入れた培養袋を、ガス透過性を有する滅菌袋に収容して滅菌袋を封止処理し、
封止処理を行った滅菌袋を滅菌ガスの雰囲気中に置いて、滅菌袋内に収容されている培養袋内の培地を滅菌処理し、
培養袋を収容した滅菌袋を封止したままの状態で、培養袋の開封部分を滅菌袋の外側から閉じ、滅菌処理を施した前記培地が入った培養袋を密封処理する、
微生物培養器の製造方法。 - 滅菌処理後、密封処理前に、滅菌処理を行った滅菌袋を真空中に置いて、滅菌袋内に残留する滅菌ガスを取り除く除去処理を更に行う、
請求項14に記載の微生物培養器の製造方法。 - 密封処理後、培養袋を減菌袋から取り出す取り出し処理を更に行う、
請求項14または15に記載の微生物培養器の製造方法。 - 培養袋の内面は、滅菌袋を形成する素材のうち少なくとも滅菌袋の内面を形成する素材よりも低い温度で熱溶着可能な素材で形成されており、
密封処理は、開封部分を滅菌袋の外側から閉じた状態で培養袋を加熱し、培養袋を密封する、
請求項14から16の何れか一項に記載の微生物培養器の製造方法。
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2016082951A (ja) * | 2014-10-29 | 2016-05-19 | Jnc株式会社 | 微生物培養器材及び微生物検出法 |
JPWO2015190090A1 (ja) * | 2014-06-09 | 2017-04-20 | 誠一 横尾 | 接着系細胞用の閉鎖系培養容器 |
EP3325601A4 (en) * | 2015-07-24 | 2019-03-13 | 3M Innovative Properties Company | THIN-LAYER CULTURE DEVICE FOR COUNTING MICRO-ORGANISMS |
JP2019062792A (ja) * | 2017-09-29 | 2019-04-25 | Jnc株式会社 | 微生物数の計測方法 |
US10808215B2 (en) | 2014-08-20 | 2020-10-20 | 3M Innovative Properties Company | Self-contained anaerobic culture device for sulfate-reducing microorganisms |
US11072771B2 (en) | 2015-09-28 | 2021-07-27 | 3M Innovative Properties Company | Self-contained anaerobic culture device with microcompartments |
US11702620B2 (en) | 2015-04-29 | 2023-07-18 | 3M Innovative Properties Company | Self-contained anaerobic environment-generating culture device |
US11851643B2 (en) | 2018-05-03 | 2023-12-26 | 3M Innovative Properties Company | Selective thin-film culture device for enumerating microorganisms |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2971094T3 (es) | 2015-04-07 | 2024-06-03 | Polyskope Labs | Detección de uno o más patógenos |
CN104911243A (zh) * | 2015-06-29 | 2015-09-16 | 汇征联合(北京)医疗器械有限公司 | 一种接种大量液体标本的固体培养基及培养方法 |
CN105886398B (zh) * | 2016-06-24 | 2019-04-09 | 郑州威瑞生物技术有限公司 | 三维内腔的软膜生物反应器及其支撑架 |
CN109790512A (zh) * | 2016-09-28 | 2019-05-21 | 捷恩智株式会社 | 测量微生物计数的方法 |
KR102235806B1 (ko) * | 2018-12-31 | 2021-04-06 | 주식회사 엔셀 | 과초산 분해 방법 및 이를 이용한 미생물의 배양방법 |
US11567092B2 (en) * | 2018-12-31 | 2023-01-31 | Robert Bosch Gmbh | System and method for operating a mold detecting device |
US11726022B2 (en) * | 2018-12-31 | 2023-08-15 | Robert Bosch Gmbh | Device for detecting mold |
WO2020195644A1 (ja) * | 2019-03-26 | 2020-10-01 | 株式会社村田製作所 | 微生物培養装置及び微生物培養方法 |
Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6142002B2 (ja) | 1979-03-30 | 1986-09-18 | Nippon Synthetic Chem Ind | |
JPH0365177A (ja) * | 1989-08-01 | 1991-03-20 | Nissho Corp | 細胞培養用バッグ |
JPH0775545A (ja) | 1993-09-09 | 1995-03-20 | Mitsubishi Gas Chem Co Inc | 嫌気性細菌簡易培養袋及びこれを用いた嫌気性細菌培養法 |
JPH08112088A (ja) | 1994-10-18 | 1996-05-07 | Business Network:Kk | 培養袋および食品の細菌検査システム |
WO1997024432A1 (fr) | 1995-12-27 | 1997-07-10 | Chisso Corporation | Dispositif d'incubation et milieu de culture de micro-organismes |
WO1998002521A1 (fr) * | 1996-07-17 | 1998-01-22 | Chisso Corporation | Milieux microbiens |
JPH11137241A (ja) | 1997-11-05 | 1999-05-25 | Otsuka Techno Kk | 培養容器 |
WO2001044437A1 (fr) | 1999-12-17 | 2001-06-21 | Chisso Corporation | Incubateur de micro-organismes et milieu de culture de micro-organismes comprenant ledit incubateur |
JP2007124985A (ja) | 2005-11-07 | 2007-05-24 | Chisso Corp | 微生物の培養方法及び微生物培養器 |
JP2008532548A (ja) * | 2005-03-14 | 2008-08-21 | コーニング インコーポレイテッド | 細胞培養フラスコ |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3552083A (en) | 1965-04-14 | 1971-01-05 | Andersen Prod H W | Apparatus and method for storing and releasing a volatile substance |
US3476506A (en) | 1965-04-14 | 1969-11-04 | Andersen Prod H W | Sterilization apparatus |
US3865695A (en) * | 1971-06-25 | 1975-02-11 | Agricole De Mycelium Du Centre | Culture of mycelium |
JP3943622B2 (ja) * | 1996-04-16 | 2007-07-11 | 株式会社日研生物医学研究所 | 細菌培養容器 |
US5736398A (en) * | 1997-01-16 | 1998-04-07 | Board Of Regents, The University Of Texas System | Gas permeable pouches for growing cultures |
US6312742B1 (en) * | 1997-02-17 | 2001-11-06 | Cryovac, Inc. | Bag-in-bag packaging system |
US6146875A (en) | 1997-05-02 | 2000-11-14 | Ward; N. Robert | Method for culturing microorganisms in prefilled flexible containers |
JP2000342246A (ja) * | 1999-06-08 | 2000-12-12 | Nissho Corp | 微生物検査用濾過容器 |
US6996952B2 (en) | 2003-09-30 | 2006-02-14 | Codman & Shurtleff, Inc. | Method for improving stability and effectivity of a drug-device combination product |
CN1942575B (zh) * | 2004-04-13 | 2012-07-18 | 东洋制罐株式会社 | 双层培养容器以及培养方法 |
US20050239200A1 (en) * | 2004-04-23 | 2005-10-27 | Beckwith Scott W | Devices for culturing anaerobic microorganisms and methods of using the same |
WO2007047028A1 (en) | 2005-10-14 | 2007-04-26 | Atrium Medical Corporation | Packaging and sterilization of medical devices |
US20070122894A1 (en) * | 2005-11-28 | 2007-05-31 | Richardson Casella Linda J | Sterile microbiological nutrient media device and methods of using |
CN201092575Y (zh) * | 2007-07-26 | 2008-07-30 | 天津百若克医药生物技术有限责任公司 | 一种分隔的细胞培养袋 |
JP5309305B2 (ja) * | 2008-01-07 | 2013-10-09 | 福岡県 | 培養バッグ及び細胞培養方法 |
-
2013
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Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS6142002B2 (ja) | 1979-03-30 | 1986-09-18 | Nippon Synthetic Chem Ind | |
JPH0365177A (ja) * | 1989-08-01 | 1991-03-20 | Nissho Corp | 細胞培養用バッグ |
JPH0775545A (ja) | 1993-09-09 | 1995-03-20 | Mitsubishi Gas Chem Co Inc | 嫌気性細菌簡易培養袋及びこれを用いた嫌気性細菌培養法 |
JPH08112088A (ja) | 1994-10-18 | 1996-05-07 | Business Network:Kk | 培養袋および食品の細菌検査システム |
WO1997024432A1 (fr) | 1995-12-27 | 1997-07-10 | Chisso Corporation | Dispositif d'incubation et milieu de culture de micro-organismes |
WO1998002521A1 (fr) * | 1996-07-17 | 1998-01-22 | Chisso Corporation | Milieux microbiens |
JPH11137241A (ja) | 1997-11-05 | 1999-05-25 | Otsuka Techno Kk | 培養容器 |
WO2001044437A1 (fr) | 1999-12-17 | 2001-06-21 | Chisso Corporation | Incubateur de micro-organismes et milieu de culture de micro-organismes comprenant ledit incubateur |
JP2008532548A (ja) * | 2005-03-14 | 2008-08-21 | コーニング インコーポレイテッド | 細胞培養フラスコ |
JP2007124985A (ja) | 2005-11-07 | 2007-05-24 | Chisso Corp | 微生物の培養方法及び微生物培養器 |
Cited By (9)
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JPWO2015190090A1 (ja) * | 2014-06-09 | 2017-04-20 | 誠一 横尾 | 接着系細胞用の閉鎖系培養容器 |
US11078455B2 (en) | 2014-06-09 | 2021-08-03 | Seiichi YOKOO | Closed culture vessel for adherent cells |
US10808215B2 (en) | 2014-08-20 | 2020-10-20 | 3M Innovative Properties Company | Self-contained anaerobic culture device for sulfate-reducing microorganisms |
JP2016082951A (ja) * | 2014-10-29 | 2016-05-19 | Jnc株式会社 | 微生物培養器材及び微生物検出法 |
US11702620B2 (en) | 2015-04-29 | 2023-07-18 | 3M Innovative Properties Company | Self-contained anaerobic environment-generating culture device |
EP3325601A4 (en) * | 2015-07-24 | 2019-03-13 | 3M Innovative Properties Company | THIN-LAYER CULTURE DEVICE FOR COUNTING MICRO-ORGANISMS |
US11072771B2 (en) | 2015-09-28 | 2021-07-27 | 3M Innovative Properties Company | Self-contained anaerobic culture device with microcompartments |
JP2019062792A (ja) * | 2017-09-29 | 2019-04-25 | Jnc株式会社 | 微生物数の計測方法 |
US11851643B2 (en) | 2018-05-03 | 2023-12-26 | 3M Innovative Properties Company | Selective thin-film culture device for enumerating microorganisms |
Also Published As
Publication number | Publication date |
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TW201428096A (zh) | 2014-07-16 |
CN104685047B (zh) | 2018-08-24 |
EP2905329A4 (en) | 2016-05-25 |
JPWO2014054494A1 (ja) | 2016-08-25 |
JP6206412B2 (ja) | 2017-10-04 |
US10030218B2 (en) | 2018-07-24 |
TWI661043B (zh) | 2019-06-01 |
US20150252314A1 (en) | 2015-09-10 |
CN104685047A (zh) | 2015-06-03 |
EP2905329A1 (en) | 2015-08-12 |
EP2905329B1 (en) | 2020-05-27 |
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