CN109652294B - Preparation and sterilization method of ready-to-use culture device - Google Patents
Preparation and sterilization method of ready-to-use culture device Download PDFInfo
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- CN109652294B CN109652294B CN201910099715.XA CN201910099715A CN109652294B CN 109652294 B CN109652294 B CN 109652294B CN 201910099715 A CN201910099715 A CN 201910099715A CN 109652294 B CN109652294 B CN 109652294B
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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
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/04—Heat
- A61L2/06—Hot gas
- A61L2/07—Steam
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/08—Radiation
- A61L2/081—Gamma radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
- A61L2/20—Gaseous substances, e.g. vapours
- A61L2/206—Ethylene oxide
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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
- C12M23/20—Material Coatings
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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
Abstract
The invention relates to the technical field of bacteriological detection for health inspection, in particular to a preparation and sterilization method for a ready-to-use culture device. In the current detection standard of coliform group, heat-resistant coliform group and most probable number of Escherichia coli (MPN) in China, a Duhan culture tube is adopted to carry out primary fermentation culture on a sample, and before the culture tube is used, air bubbles in a built-in small inverted tube need to be completely discharged, so that the operation is time-consuming and labor-consuming. The application provides an external culture tube of trachea, greatly reduced the degree of difficulty of exhausting before using, the producer can directly fill the culture medium preparation and become the culture tube of a ready-to-use type, this application provides a preparation method and sterilization method to this ready-to-use type culture tube, adopts the sterilization method in this application, can once only accomplish the sterilization of big batch product, practices thrift the sterilization step, and it is poor to improve production efficiency.
Description
Technical Field
The invention relates to the technical field of hygiene test bacteriological detection, in particular to a preparation and sterilization method of a ready-to-use culture device for detecting coliform group bacteria, heat-resistant coliform group bacteria and Most Probable Number (MPN) of Escherichia coli.
Background
Detection of coliform group, heat-resistant coliform group (faecal coliform group) and Escherichia coli in hygienic microbiological indicators of public products such as water, food, cosmetics and public places includes multitubular fermentation Most Probable Number (MPN) method, filtration membrane method, plate counting method, enzyme substrate method and the like. The multi-tube fermentation Most Probable Number (MPN) method is widely applied by the advantages of low technical difficulty, low reagent cost, wide detection range and the like, and particularly has outstanding advantages in small-scale laboratory application. In the national GB/T5750.12-2006 Standard test method for domestic Drinking Water microbial indicators, the test of Total coliform, Heat-resistant coliform and Escherichia coli, the test of Escherichia coli count in GB 4789.3-2016 food safety national Standard food microbiology, the test of Escherichia coli count in GB 4789.38-2012 food safety national Standard food microbiology, the test of Escherichia coli count in GB/T18204.10-2000 swimming pool Water microbial test method, and the test of Multi-tube fermentation Most Probable Number (MPN) methods, such as the test of Total coliform in Water and the test of fecal coliform in Water and wastewater monitoring and analysis method (fourth edition), are the first methods; in the inspection of faecal coliform in technical specifications for cosmetic safety (2015 edition), the inspection of faecal coliform population in GB 4789.39-2013 food safety national standard food microbiology, and the inspection of hygiene in GB/T18204.4-2013 public places fourth part: a multi-tube fermentation method of coliform group in public appliance microorganism, namely, a multi-tube fermentation Most Probable Number (MPN) method in an inspection method of fecal coliform group in sewage and sludge in medical institution, namely, appendix A (normative appendix) of GB-18466 and 2005 medical institution water pollutant discharge standard of medical institution, and the like are only methods.
The basic experimental process of the multitube fermentation Most Probable Number (MPN) method comprises primary fermentation, plate separation and secondary fermentation, and the acid and gas production phenomenon is observed after the coliform group is cultured for a period of time on the basis of the biochemical characteristic that the coliform group can ferment lactose to produce acid and gas. In the existing detection process, a test tube containing a lactose culture medium (such as lactose peptone culture medium, lactose cholate fermentation culture medium (containing a neutralizing agent), brilliant green lactose cholate broth, lauryl sulfate tryptone broth (LST), lactose broth, and currently-hidden broth) with a built-in small inverted tube (inverted Duhan tube) is generally adopted to culture a sample to be tested, the effect of acid and gas production is observed, the test tube used for culture needs to be self-made in a front laboratory, namely the laboratory prepares the culture medium by itself, the Duhan tube is inverted and placed into the test tube, the culture medium is subpackaged, high-pressure sterilization and degassing are carried out, the Duhan tube is washed after culture, and the like, the process is complex, and a large amount of manpower is consumed. The technical bottleneck restricting instant commercialization of the culture tube lies in that the Duhan tube used at present is as shown in figure 1, a small test tube is inverted in a slender culture tube, if a finished product culture tube prepared by a manufacturer is used, air easily enters the small inverted tube after vibration such as transportation, the caliber of the small inverted tube is smaller and forms an inverted state with a 180-degree included angle with a fermentation test tube, gas is difficult to discharge before use, a proper angle needs to be selected by a skilled experimenter, liquid is slowly filled, gas is pressed out, each tube needs to be operated carefully, and long time is needed. The inventor provides a ready-to-use culture device for detecting flora by a most probable method in order to solve the bottleneck technology, and the ready-to-use culture device adopts an external gas production pipe, is connected with a main pipe through a bent connecting pipe, increases the selection freedom of the inner diameter and the inversion angle of the gas production pipe, and can obviously reduce the difficulty of removing bubbles in the gas production pipe.
Disclosure of Invention
The culture tube is provided with a hard main tube and a hard gas production tube, which are connected through a bent connection tube, and the closed hard gas production tube is adopted to replace a built-in small inverted tube in the prior art, is not arranged in the fermentation tube any more and is not limited by the built-in tube diameter and the inverted position any more; thereby the inner diameter and the inversion angle of the hard gas production collecting pipe are increased in freedom degree, the pipeline connection between the hard main pipe for fermentation and the hard gas production collecting pipe is smoother, and the gas is convenient to discharge. The culture tube is applied to industrial production, a manufacturer can put in a dry powder culture medium after production is finished and then seal and package the culture tube, and technicians can use the culture tube only by opening the package and adding water of a proper solvent when using the culture tube, so that the culture tube is quick, efficient, time-saving and labor-saving.
Aiming at the technical scheme, the application further provides a preparation method and a sterilization method of the ready-to-use fermentation culture tube, and the sterilization method in the application can realize one-time large-batch sterilization and save production steps.
In order to achieve the above technical effects, the present application provides the following technical solutions:
in a first aspect of the present application, a culture tube is provided, which comprises a main tube, a bent tube and an air-producing tube; the main pipe is a cylindrical member, the elbow pipe is a cylindrical member bent at a certain angle, the gas production pipe is a cylindrical member with one closed end, the main pipe, the elbow pipe and the gas production pipe are sequentially connected end to end, the length of the main pipe is greater than that of the gas production pipe, and the inner diameter of the main pipe is greater than that of the gas production pipe.
Preferably, the main tube and the gas production tube are made of hard materials.
Preferably, the main tube and the gas production tube are made of one of synthetic resin, organic glass, high borosilicate glass and polypropylene.
Preferably, the gas generating tube includes an extension portion and a sealing portion, the extension portion is a cylindrical member, and the sealing portion and the extension portion are integrally formed to constitute a closed cavity.
Further preferably, the sealing portion has a hemispherical configuration protruding outward along the extension portion.
Further preferably, the sealing portion is a sheet-like portion that closes one end of the extending portion.
Preferably, the gas production pipe comprises an extension part and a sealing part, the extension part is a cylindrical member, and the sealing part is a detachable plug.
Further preferably, the plug is made of an air-impermeable material.
Preferably, the elbow is made of soft materials. Further preferably, the elbow is a soft cavity pipeline made of silica gel, rubber or plastic.
Preferably, the main pipe, the elbow pipe and the gas production pipe are integrally formed to form a section of continuous cavity with one end being closed.
Preferably, the bending angle of the elbow is 35 to 180 degrees.
Preferably, the inner diameter of the main tube and the gas production tube is 0.6-4cm, and the length is 3-25 cm.
Preferably, the culture tube is further provided with a tube cap, and the size of the tube cap is matched with that of the main tube and used for covering the main tube.
Further preferably, the pipe cap is made of water-proof and breathable silica gel, rubber or plastic materials.
In a second aspect of the present application, a ready-to-use culture tube for use in an MPN detection method is prepared by adding a culture medium to the culture tube.
Through the description to above-mentioned culture tube structure, the culture tube in this application has continuous cavity and necking down structure, and the person in charge and produce the trachea and be certain contained angle, and the sample gathers in producing the trachea top and form a section macroscopic cavity in the gas that the fermentation in-process produced to accomplish the process of detecting. The culture tube of this structure can show and reduce the exhaust degree of difficulty, and the producer can fill the culture medium into the pipe after accomplishing the production of culture tube, prepares into one kind and tears the culture tube that the packing can be used.
In a third aspect of the present application, a method for preparing the above culture tube is provided, where the method for preparing the culture tube includes a split culture tube preparation method and an integrated culture tube preparation method:
the split culture tube is used for preparing a main tube and an air production tube through injection molding or blow molding, and the bent connection tube is prepared through a mould pressing forming method by adopting an organic silica gel material;
preferably, the elbow has a hardness of 40-70 degrees.
Aiming at the split type culture tube, a separated main tube, a bent connection tube and an air production tube are connected in a splicing mode. The research of the application finds that the elbow pipe prepared by adopting the organic silicon material is non-toxic and tasteless, has stable chemical performance and mechanical strength, and the adsorbability of the organic silicon material can ensure that the split culture pipe is more firmly connected.
The preparation method of the integrated culture tube comprises a preparation method of the culture tube with an integrated gas production tube and a preparation method of the culture tube with a plug at the sealing part of the gas production tube;
the culture tube with the integrated gas production tube is prepared by a blow molding method, a mold with a corresponding specification is selected, and demolding is carried out after blow molding;
the research of the application finds that the culture tube with the structure has better sealing part forming effect compared with injection molding by adopting the blow molding mode,
the culture tube with the gas production tube sealing part as the plug is prepared in an injection molding mode, and is subjected to core-pulling and demolding after injection molding through a mold. Thermoplastic resin raw materials are injected into the split mold, the pipe wall is attached to the inner wall of the mold through introducing compressed air, the sealing part with the corresponding structure can be obtained through controlling air flow, and the cost of opening the mold in a factory is saved.
Preferably, the inner diameter of the main pipe of the culture tube is 0.6-4cm, the length of the main pipe is 3-25cm, the inner diameter of the gas production tube is 0.6-4cm, and the length of the gas production tube is 3-10 cm; the included angle degree of the bent pipe part is 0-135 degrees, the diameter of the inner cavity at the joint with the main pipe is 0.7-4.1cm, the diameter of the inner cavity at the joint with the gas production pipe is 0.7-4.1cm, and the wall thickness is 0.1-0.5 cm.
According to national standard detection standard, usually the culture medium volume in a culture tube is about 10ml, and the culture tube in this application is responsible for and is appeared certain contained angle with producing the trachea, and need be with the gaseous evacuation in the producing the trachea before the use, adopts the culture tube of above-mentioned size can guarantee that the culture medium in the culture tube reaches effective liquid level height, accomplishes and detects.
Preferably, the culture tube is added with dry powder or liquid culture medium, then a tube cap is covered, and the culture tube is placed in a packaging bag for sealing.
Preferably, the culture tube added with the liquid culture medium is a culture tube made of polypropylene.
In a fourth aspect of the present application, a method for sterilizing the ready-to-use culture tube is provided, wherein a plug and a cap are added to the culture tube containing the dry powder culture medium, the culture tube is packed in an independent packaging bag, the culture tube is sterilized by gamma rays in an unsealed state, and the outer packaging is sterilized by ethylene oxide after the sealing;
or preparing corresponding single-material, double-material or three-material culture medium liquid culture medium according to a sample detection standard, adding the culture medium liquid culture medium into a culture tube made of polypropylene, adding a plug made of polypropylene and a silicon rubber tube cap, sterilizing the culture medium liquid culture medium by high-temperature steam, filling the culture medium liquid culture medium into an independent packaging bag, sealing the independent packaging bag, and sterilizing the external packaging bag by adopting ethylene oxide.
Under the normal condition, different parts have different requirements on sterilization conditions, manufacturers need to sterilize the culture tubes in the preparation process and then assemble the culture tubes, and products are possibly infected with bacteria again in the assembly process, so the sterilization process needs to be repeated before packaging and after sealing.
Because the liquid culture medium is not suitable for ethylene oxide sterilization and radiation sterilization, and high-pressure steam sterilization is needed, the culture tube of the liquid culture medium is prepared by adopting the high-pressure-resistant polypropylene material, the liquid culture medium can be directly filled into the culture tube, the whole culture medium is sterilized by high-pressure steam, and the sterilization steps are saved.
The culture tube is sealed after being filled with the dry powder culture medium, and is sterilized by gamma rays in an unsealed state, so that the culture tube in the dry powder state can achieve a good penetrating effect, and the sterilization effect is thorough. Because package seal adopts hot melt or sticky mode usually, this application research discovery adopts gamma ray to disinfect to outer packing and can influence the sealed effect of packing, and ethylene oxide can disinfect under the low temperature condition, sterilizes this article extranal packing, and sterilization effect is good and can not influence the quality of packing.
Preferably, the dose of the gamma ray is 17-19 kGy, and more preferably, 18 kGy.
The gamma ray irradiation with the dose can instantly finish the sterilization process, can well penetrate through the tube wall to sterilize the inside, and can not influence the packaging quality. Low doses make it difficult to sterilize thoroughly, and high doses may cause damage to the product itself.
Preferably, the ethylene oxide sterilization concentration is 700-800 mg/L, the temperature is 55-60 ℃, the relative humidity is 60-80%, and the action time is 3-4 h.
The ethylene oxide sterilization usually adopts the form of sterilization storehouse, and to the ready-to-use culture tube in this application, adopt above-mentioned parameter to carry out ethylene oxide sterilization and can realize that the ready-to-use culture tube in a unit sterilization storehouse obtains thorough sterilization effect. Conditions of excessive temperature and humidity reduce the seal, while conditions of too low temperature are detrimental to complete sterilization of the entire package.
Preferably, the independent packaging material is low-density polyethylene, high-density polyethylene, polypropylene, polyvinyl chloride or polystyrene.
The research of the application shows that the package prepared by the materials can meet the requirement of good sealing performance.
Preferably, the liquid culture medium is sterilized by high-temperature steam at 121 ℃ for 15 min.
The invention has the advantages of
The application provides a culture tube with an external gas production tube, overcomes the technical defect that the exhaust is difficult before the use of a Duhan tube in the prior art, provides a preparation method and a sterilization method of the culture tube, provides corresponding preparation and sterilization methods for different forms of culture tubes in the application, and ensures that products prepared according to the sterilization method in the application meet the gamma-ray irradiation sterilization standard of GB16352-1996 disposable medical supplies, and can obviously save production steps when applied to industrial production.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the application and, together with the description, serve to explain the application and are not intended to limit the application.
FIG. 1 is a schematic view of a Duhan's fermentation tube used in the prior art;
FIG. 2 is a ready-to-use culture tube in example 1;
wherein 11 is a cap, 12 is a main tube, 13 is an elbow tube, 14 is an air generating tube, 141 is a sealing part, 142 is an extension part, and A1 is a culture solution in a use state.
FIG. 3 is a ready-to-use culture tube in example 2;
wherein 11 is a cap, 12 is a main tube, 13 is a bent tube, 15 is an air-producing tube, 151 is an extension part, 152 is a sealing part, and A1 is a culture solution in use.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
In the current coliform group detection standard in China, a Duhan's culture tube is adopted to carry out primary fermentation culture on a sample, and before the culture tube is used, bubbles in a built-in small inverted tube need to be completely discharged, so that the operation is time-consuming and labor-consuming. The application provides an external culture tube of trachea, greatly reduced the degree of difficulty of exhausting before using, the producer can directly fill the culture medium preparation and become the culture tube of a ready-to-use type, this application provides a preparation method and sterilization method to this culture tube of ready-to-use type.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific examples and comparative examples.
Example 1
The culture tube in this embodiment is shown in FIG. 2, and comprises a main tube 12, an elbow tube 13 and a gas-producing tube 14; the main pipe 12 is a cylindrical member, the elbow pipe 13 is a cylindrical member bent at a certain angle, the gas production pipe 14 is a cylindrical member with one end closed, and the main pipe 12, the elbow pipe 13 and the gas production pipe 14 are sequentially connected and integrally formed to form a section of continuous cavity with one end closed.
The main tube 12 and the gas generating tube 14 are made of synthetic resin.
The gas generating tube 14 includes an extension portion 142 and a sealing portion 141, the extension portion 142 is a cylindrical member, and the sealing portion 141 and the extension portion 142 are integrally formed to constitute a closed cavity.
The sealing portion 141 has a hemispherical configuration protruding outward along the extension.
The bending angle of the elbow pipe is 60 degrees.
The main tube 12 and the gas-producing tube 14 have inner diameters of 2cm and 0.6cm, respectively, and lengths of 20cm and 10cm, respectively.
The culture tube is also provided with a tube cap 11, and the tube cap 11 is covered on the main tube 12.
The pipe cap 11 is made of waterproof and breathable silica gel.
A1 is the medium in use.
Example 2
The culture tubes in this example are shown in FIG. 3: the culture tube comprises a main tube 12, an elbow tube 13 and an air production tube 15; the main pipe 12 is a cylindrical member, the elbow pipe 13 is a cylindrical member bent at a certain angle, the gas production pipe 15 is a cylindrical member with one end closed, and the main pipe, the elbow pipe and the gas production pipe are sequentially connected.
The main tube 12 and the gas generating tube 15 are made of synthetic resin.
The gas production tube 15 comprises an extension part 151 and a sealing part 152, the extension part 151 is a cylindrical component, the sealing part 152 is a detachable plug, and the plug is made of an airtight material.
The bending angle of the elbow pipe 13 is 120 degrees.
The inner diameters of the main pipe 12 and the gas production pipe 15 are respectively 3cm and 1cm, and the lengths thereof are respectively 23cm and 5 cm.
The culture tube also has a tube cap 11 for covering the main tube. The pipe cap 11 is made of plastic.
A1 is the medium in use.
Example 3
The embodiment provides a preparation method and a sterilization method of a split culture tube, wherein a main tube, an air production tube and a bent connection tube mold are respectively opened, the main tube and the air production tube are prepared by injection molding, and the bent connection tube is prepared by adopting an organic silica gel material through a die pressing forming method; the elbow hardness is 50 degrees. Then the main pipe, the elbow pipe and the gas production pipe are spliced and assembled in sequence to form the culture pipe.
Adding a lactose bile salt fermentation dry powder culture medium into the spliced culture tube, covering a tube cap, filling the tube cap into a low-density polyethylene independent packaging bag, sterilizing by gamma rays of 18 kGy, sealing the packaging bag by hot melting, and then putting the packaging bag into an ethylene oxide sterilization bin to sterilize the external packaging; the sterilization concentration of the ethylene oxide is 750mg/L, the temperature is 58 ℃, the relative humidity is 75 percent, and the action time is 3.5 h.
Example 4
In this embodiment, a method for manufacturing an integrated culture tube and a method for sterilizing the same are provided, in which a mold is first opened, and a polypropylene thermoplastic material is blow molded and then demolded. Filling the prepared polypropylene culture tube with a double-material lactose bile salt fermentation liquid culture medium with a use concentration, adding a polypropylene tube cap, and sterilizing the liquid culture medium in a high-temperature steam sterilization pot under the high-temperature steam sterilization condition of 121 ℃ for 15 min.
And (3) putting the sterilized culture tube into an independent packaging bag for sealing, and putting the sealed packaging bag into an ethylene oxide sterilization bin for sterilization, wherein the ethylene oxide sterilization concentration is 800mg/L, the temperature is 60 ℃, the relative humidity is 80%, and the action time is 4 h.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (9)
1. A ready-to-use culture tube applied to an MPN detection method is characterized by comprising a main tube, an elbow tube and an air production tube; the main pipe is a cylindrical member, the elbow pipe is a cylindrical member bent at a certain angle, the gas production pipe is a cylindrical member with one closed end, the main pipe, the elbow pipe and the gas production pipe are sequentially connected end to end, the length of the main pipe is greater than that of the gas production pipe, and the inner diameter of the main pipe is greater than that of the gas production pipe;
the culture tube is also provided with a tube cap, and the size of the tube cap is matched with that of the main tube and used for covering the main tube; the pipe cap is made of waterproof and breathable silica gel, rubber or plastic;
the gas production pipe comprises an extension part and a sealing part, the extension part is a cylindrical component, and the sealing part and the extension part are integrally formed to form a closed cavity;
the sealing part is of a hemispherical structure protruding outwards along the extension part or is of a sheet shape and seals one end of the extension part;
the bending angle of the bent connecting pipe is 35-180 degrees; the inner diameters of the main pipe and the gas production pipe are 0.6-4cm, and the lengths of the main pipe and the gas production pipe are 3-25 cm;
the culture tube also has a culture medium therein.
2. The ready-to-use culture tube for use in the MPN detection method according to claim 1, wherein the gas production tube comprises an extension part and a sealing part, the extension part is a cylindrical member, and the sealing part is a detachable plug; the plug is made of an airtight material.
3. The ready-to-use culture tube for MPN detection method according to claim 1, wherein the elbow is a soft hollow tube made of silica gel, rubber or plastic; the main pipe, the elbow pipe and the gas production pipe are integrally formed to form a section of continuous cavity with one end being closed.
4. The method for preparing a ready-to-use culture tube for use in the MPN detection method according to any one of claims 1 to 3, wherein the preparation method comprises a split culture tube preparation method and an integrated culture tube preparation method:
the split culture tube is used for preparing a main tube and an air production tube through injection molding or blow molding, and the bent connection tube is prepared through a mould pressing forming method by adopting an organic silica gel material;
the preparation method of the integrated culture tube comprises a preparation method of the culture tube with an integrated gas production tube and a preparation method of the culture tube with a plug at the gas production tube sealing part:
the culture tube with the integrated gas production tube is prepared by a blow molding method, a mold with a corresponding specification is selected, and demolding is carried out after blow molding;
the culture tube with the gas production tube sealing part as the plug is prepared in an injection molding mode, and is subjected to core-pulling and demolding after injection molding through a mold.
5. The method according to claim 4, wherein the inner diameter is 0.6-4cm, the length is 3-25cm, the inner diameter of the gas-producing tube is 0.6-4cm, the length is 3-10cm, the included angle of the elbow is 0-135 degrees, the diameter of the inner cavity at the connection with the main tube is 0.7-4.1cm, the diameter of the inner cavity at the connection with the gas-producing tube is 0.7-4.1cm, and the wall thickness is 0.1-0.5 cm.
6. The sterilization method for a ready-to-use culture tube for MPN detection method according to any one of claims 1 to 3, wherein the culture tube filled with the dry powder medium is capped with a stopper and a cap, packed in a separate packaging bag, sterilized by gamma ray in an unsealed state, and sterilized by ethylene oxide after sealing;
or preparing corresponding single-material, double-material or three-material culture medium liquid culture medium according to a sample detection standard, adding the culture medium liquid culture medium into a culture tube made of polypropylene, adding a plug made of polypropylene and a silicon rubber tube cap, sterilizing the culture medium liquid culture medium by high-temperature steam, filling the culture medium liquid culture medium into an independent packaging bag, sealing the independent packaging bag, and sterilizing the external packaging bag by adopting ethylene oxide.
7. The method for sterilizing a ready-to-use culture tube for an MPN detection method according to claim 6, wherein the gamma ray dose is 17-19 kGy.
8. The sterilization method for the ready-to-use culture tube for the MPN detection method according to claim 6, wherein the dose of the gamma ray is 18 kGy; the ethylene oxide sterilization concentration is 700-800 mg/L, the temperature is 55-60 ℃, the relative humidity is 60-80%, and the action time is 3-4 h.
9. The sterilization method for the ready-to-use culture tube for the MPN detection method according to claim 6, wherein the individual packaging material is low density polyethylene, high density polyethylene, polypropylene, polyvinyl chloride, polystyrene; the liquid culture medium is sterilized by high-temperature steam at 121 deg.C for 15 min.
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| CN206828536U (en) * | 2017-05-22 | 2018-01-02 | 华南理工大学 | A kind of coliform detection means |
| CN108179171A (en) * | 2017-12-12 | 2018-06-19 | 天津科技大学 | A kind of method of aerogen in Rapid identification swollen bag soy sauce |
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| US20040166556A1 (en) * | 2003-02-21 | 2004-08-26 | Cullimore Dennis Roy | Analytic device for the detection and quantification of fermentative cultured activities |
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| US5108701A (en) * | 1989-05-15 | 1992-04-28 | Cem Corporation | Process for rapid sterilization of biological media |
| CN101029334A (en) * | 2006-11-30 | 2007-09-05 | 白莉 | Colicine testing fluid and simple testing bottle |
| CN104098915A (en) * | 2014-07-23 | 2014-10-15 | 赵雪雅 | Modified silica gel material, and preparation method and application thereof |
| CN105997156A (en) * | 2016-06-12 | 2016-10-12 | 杭州昕龙医疗科技有限公司 | Sampling swab for full-automatic pretreatment |
| CN206828536U (en) * | 2017-05-22 | 2018-01-02 | 华南理工大学 | A kind of coliform detection means |
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