JP2000262269A - Culture device - Google Patents

Abstract

(57) [Summary] [PROBLEMS] To provide a culture apparatus that can perform a series of culture operations such as sampling, introduction of a culture medium, and culture with one apparatus. SOLUTION: An elastic porous body for supporting a culture medium and a case for holding the porous body therein are provided, and a part of the case has an openable and closable opening, and the presence or absence of microorganisms from this opening. A culture apparatus configured to take in and out a sample or a medium to be used for the test.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a microorganism culturing apparatus.

[0002]

2. Description of the Related Art Generally, a liquid submerged culture method is used for culturing microorganisms. Liquid submerged culture is a culture method in which nutrients and dissolved oxygen are efficiently supplied by shaking a culture flask. In clinical tests, an agar culture method is often used.

[0003]

The liquid submerged culture method comprises:
Means such as shaking and stirring are required, and the apparatus using this culture method is large and complicated. Furthermore, since the liquid medium is handled during the injection and extraction of the bacterial solution sample, the operation requires careful operation. In addition, when a liquid containing microorganisms is moved, there is a risk that the microorganisms may contaminate the handler and the environment. On the other hand, in the agar culture method, a shaking device is not required, but the contact between the culture medium and the microorganism is flat, and in order to favor the growth of the microorganism, when adding the microorganism, an operation of spreading and spreading thinly is required. is necessary. A skilled technique is required for both liquid submerged culture and agar culture.

[0004]

SUMMARY OF THE INVENTION In order to solve the above problems, the present invention comprises an elastic porous body for supporting a culture medium and a case for holding the porous body inside, and a part of the case. The present invention provides a culture apparatus that has an opening that can be opened and closed, and that allows a sample or a culture medium to be used for testing for the presence or absence of microorganisms to be taken in and out of the opening. The culture apparatus of the present invention disperses microorganisms in a porous body supporting a medium containing a component effective for the growth of microorganisms and cultures the microorganisms in the porous body, and the medium completely fills the pores of the porous body. Is carried in an amount that cannot be satisfied.
By adopting this method, the exchange between the sample medium and the liquid medium can be performed with one device, and a culture that does not require a shaking device, requires no skill, and further reduces the risk of contamination. It becomes possible.

Further, the present invention employs a system in which a sample or a medium is taken in and out from an opening provided in a part of a case. This can reduce the risk of contamination when handling the bacterial solution sample. Furthermore, by providing a region for identifying the culture state of the microorganism in the case, the proliferation degree can be assayed without extracting the bacterial solution. If the medium impregnated in the porous body is in a powder or liquid form, a suitable environment for microorganisms can be easily provided in the apparatus.

It is preferable that the case has means for pushing out the contents contained in the porous body or the porous body itself out of the case, or sucking the contents from the outside of the case. This makes it possible to easily and safely obtain the bacterial solution after culturing, and to provide it for inspection work. Further, it is preferable that a medicine having a bactericidal action is held in the case, and a means is provided for killing the microorganism after culture by bringing the medicine into contact with the porous body. Thereby, the operation can be made more secure, and the concern of environmental pollution after use can be prevented.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION The porous material used in the present invention has a material type, material, or pore size which satisfies the necessary and sufficient conditions for culturing by dispersing microorganisms.
It is made of elastic material. The types of materials constituting the porous body vary widely. Assuming that the definition of porous body has pores, a foam having three-dimensionally connected pores,
For example, a sponge-like material, a woven or non-woven fabric of elastic fibers having a capillary effect.

As the material of the porous body, a material having high hydrophilicity is effective because the culture solution of the microorganism is an aqueous solution, and examples thereof include cellulose or a derivative thereof, and glass. Further, even in the case of non-hydrophilic plastics, metals, and the like, materials whose surfaces are subjected to chemical treatment or plasma treatment to impart hydrophilicity are also effective. The pore diameter of the pores of the porous body satisfies at least a necessary and sufficient condition for dispersing microorganisms. Specifically, when the target is Escherichia coli, a structure having pores having a pore diameter in the range of 10 μm to several mm is desirable. The porous body having elasticity is compressed by an external force of a piston or the like in the case, and can easily return to its original state when the external force is released. is there.

As the material of the porous body, a high molecular polysaccharide, a polypeptide or a complex of both is preferable. Among high molecular polysaccharides, cellulose, amylose or amylopectin are preferred. The cellulose preferably has a functional group such as a hydroxyl group, an alkoxyl group, an acyl group, or an amino group provided by a chemical treatment.

According to the present invention, by using a stretchable elastic porous body, it is possible to extract only the medium after sampling the sample, and subsequently to inhale the liquid medium, and to exchange only the liquid. Become. For this purpose, it is preferable that the inside of the porous body has such a property that water moves at a speed higher than that of microorganisms. A medium that provides an effective environment for the growth of microorganisms is, for example, a medium used in pre-enrichment culture. The medium that promotes or suppresses the growth of microorganisms is, for example, a medium used in separation culture. In one embodiment, the elastic porous body is enclosed in a case made of, for example, plastics, has a structure in which there is no risk of infection to a user in a culture process, microorganisms do not go outside, and The structure does not contain foreign matter.

In another embodiment, the structure is such that sampling of a sample can be easily performed. That is, the case has an openable / closable opening so that the sample can be impregnated into the elastic porous body in the case from the opening. Alternatively, the elastic porous body is exposed at the opening, and the sampling can be performed by immersing the exposed part in the sample or wiping the sample attached to the holder.

In still another embodiment, an elastic porous body impregnated with a medium and an elastic porous body for culture are placed in a case so that a sample or a medium can be circulated between the porous bodies. I do. Then, by providing a filter in a part of the circulation path and circulating the sample or medium,
A large contaminant higher than the microorganism in the sample is removed by a filter.

In another embodiment, the case is provided with a transparent region so that the porous body can be observed from the outside. When identifying the growth or detection of microorganisms, the bacterial solution after culturing or the elastic porous body containing the bacterial solution can be identified in a transparent region of the case without being taken out of the case. Alternatively, an openable door is provided in the case, and when the door is opened, the elastic porous body can be seen. Further, in order to facilitate identification of the growth or detection of the microorganism, a porous body containing a reagent capable of easily determining the growth of the microorganism, for example, a luminescent reagent or a staining reagent, is separately added. Alternatively, change the color of the elastic porous body
A color that facilitates identification of the growth or detection of microorganisms. In another embodiment, not only to identify the inside of the case, but also to directly take out the cultured bacterial solution or the elastic porous body containing the same from there,
The area for identification is openable.

In another embodiment, after the sample is sampled, the powdered medium is dispersed in the elastic porous body so that the inside of the elastic porous body in the case can be efficiently made into an environment suitable for the growth of microorganisms. I have. Alternatively, the liquid medium is in a state of permeating the inside of the elastic porous body.
Alternatively, a capsule packed with a powdered medium is inside the resilient porous body. The capsule here is, for example,
It is a capsule-like drug used for pharmaceuticals and the like, and the material is preferably a water-soluble one. The size or number of capsules is large enough to fit inside the elastic porous body to be cultured.When water is contained in the elastic porous body, the capsule dissolves and the elastic porous body is impregnated with the medium. Size or quantity that satisfies the necessary and sufficient conditions for

[0015] In still another embodiment, the case has a structure capable of extracting the bacterial solution after being cultured in the porous body inside the case, for example, a structure like a syringe. In another embodiment, the post-cultivation treatment includes:
It has a structure that can kill microorganisms with drugs. For example, by incorporating a porous body impregnated with a surfactant and bringing it into contact with the elastic porous body after cultivation, a structure is obtained in which the surfactant can be sent to the elastic porous body cultured with microorganisms. ing. Alternatively, there is a surfactant in a glass sealed tube, and the structure is such that the elastic porous body after culturing can be immersed in the surfactant by breaking the glass sealed tube. In another embodiment, a function for controlling the temperature of the culture is attached, for example, a Peltier device is contained in a case.

Next, some embodiments will be described with reference to the drawings. The culturing apparatus shown in FIG. 1 includes a plastic syringe type case 1 and a cellulose elastic porous body 2 placed under the case. The elastic porous body 2 is impregnated with a medium. In the case 1, a piston 3 with a rubber plate 6 is inserted above the elastic porous body 2. A sample injection port 4 is provided at the center of the piston 3, and the injection port is sealed with a cap 5 except when a sample is injected. At the bottom of the case 1, a sample extraction port 7 is provided.
This extraction port is always sealed with a cap 8. In order to culture microorganisms using this apparatus, dry heat sterilization is performed in advance. Next, the cap 5 is opened and the injection port 4
, A microbe is introduced into the elastic porous body 2, and is cultivated under culturing conditions, for example, a predetermined temperature. After the cultivation, the elastic porous body 2 is compressed by the piston 3 to extract the proliferated microorganisms from the extraction port 7.

The culturing apparatus shown in FIG. 2 has a case 11 having an opening at the lower end and a resilient porous body 12 housed with its tip protruding outside through the opening. Sampling can be performed by immersing the tip of the elastic porous body in the sample or wiping the sample attached to the holder. After sampling the sample in this manner, at the tip of the case 11,
A cap 13 large enough to cover the protruding portion of the elastic porous body is fitted. The sample is cultured in this state.

The culturing apparatus shown in FIG. 3 is a combination of a case 21 containing an elastic porous body 25 for culture and a case 29 containing an elastic porous body 28 carrying a medium or a bactericide. The case 21 includes a piston 22 having a sample inlet 31, a rubber plate 24 attached to a lower end thereof,
An extraction port 26 for extracting a sample after culturing is provided. On the other hand, a piston 23 having a rubber plate 30 is fitted into the case 29, and a nozzle 27 is provided at the tip.
The nozzle 23 is inserted into an opening 32 provided in the case 21 and is combined as shown in the drawing.
The medium or the bactericide can be injected into the medium. That is, the medium or the bactericide impregnated in the elastic porous body 28 compresses the porous body 28 with the piston 23,
It is added from the nozzle 27 to the elastic porous body 25 in the case 21.

The culture apparatus shown in FIG.
A partition wall 46 having fine pores
This is an example in which a case 42 accommodating a resilient porous body 45 is combined. A rubber plate 44 and a piston 41 are inserted into the case 42, and a medium or a bactericide impregnated in the porous body 45 is appropriately passed through the pores of the partition wall 46 to the elastic porous body 47 by operating the piston 41. Can be added. Reference numeral 48 denotes an extraction port, which is always sealed with a cap.

[0020]

EXAMPLE An example in which a growth curve equivalent to that obtained by liquid shaking culture was obtained in a culture test using the culture apparatus of the present invention will be described below. The culture apparatus having the structure shown in FIG. 1 was used. Cellulose sponge was used for the porous body. This was put in the lower part of the plastic syringe type case 1. The porous body 2 was impregnated with a medium. After the apparatus was dry-heat sterilized in advance, the porous body 2 was put therein, the cap 5 was opened, and an Escherichia coli solution was put through the inlet 4 to impregnate the porous body 2. In this state, E. coli was cultured at 37 ° C. Eight of these culture devices were prepared, and each of them was 1, 2, 3, 4, 5, 6, 7,
Alternatively, after 8 hours, the Escherichia coli solution after the growth by compressing the porous body 2 with the piston 3 was extracted from the extraction port 7. FIG. 5 shows the concentration of E. coli for each elapsed time.

[0021]

As described above, according to the present invention, it is possible to obtain a culturing apparatus having the same culturing ability as that of liquid shaking culture and more efficient culturing ability than agar culture. In addition, by providing an additional function such as a combination of a resilient porous body and a case and a piston, a series of operations in culture such as sampling, culture, and extraction can be easily performed. Therefore, there is no danger such as contamination due to the use of the culture device, and it becomes a general-purpose culture device, greatly contributing to the field of microbial research or microbial testing, and also applicable to household testing systems. Become.

[Brief description of the drawings]

FIG. 1 is a longitudinal sectional view of a syringe-type culture apparatus according to one embodiment of the present invention.

FIG. 2 is a longitudinal sectional view of a culture apparatus according to another embodiment of the present invention.

FIG. 3 is a longitudinal sectional view of a culture apparatus according to still another embodiment of the present invention.

FIG. 4 is a longitudinal sectional view of a culture apparatus according to still another embodiment of the present invention.

FIG. 5 is a diagram comparing a growth curve using a syringe-type culture device of the present invention with a growth curve obtained by a liquid shaking culture method.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Case 2 Porous body 3 Piston 4 Injection port 5 Cap 6 Rubber plate 7 Extraction port 8 Cap

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Hitoshi Miyazaki 1006 Kazuma Kadoma, Kadoma City, Osaka Prefecture F-term in Matsushita Electric Industrial Co., Ltd. 4B029 AA02 AA08 AA21 BB02 CC10 DF05 DG06 DG08 GA03 GB01 GB05 GB07 GB10

Claims (10)

[Claims] 1. A resilient porous body for supporting a culture medium and a case for holding the porous body therein, and a part of the case has an openable and closable opening, and the opening of the microorganism is A culture device configured to take in and out a sample or medium to be tested for the presence or absence. 2. The culture apparatus according to claim 1, wherein the inside of the porous body has such a property that water moves at a speed higher than that of microorganisms. 3. An environment in which microorganisms can be exchanged by capturing only microorganisms contained in an aqueous sample by expanding and contracting the porous body and separately sucking a medium effective for the growth of the microorganisms. The culture device according to claim 1, wherein the culture device is configured as follows. 4. The culture apparatus according to claim 1, wherein the culture medium contains a component that promotes or suppresses the growth of a specific microorganism. 5. The culture apparatus according to claim 1, wherein a porous body is arranged so as to face the opening of the case, and a sample and a medium are taken in and out through the porous body. 6. The culture apparatus according to claim 1, wherein the case includes an area having means for identifying a culture state of the microorganism. 7. The culture apparatus according to claim 1, wherein the porous body or a medium carried on the porous body is impregnated with a reagent for identifying the degree of growth of microorganisms. 8. The culture apparatus according to claim 1, further comprising at least two porous bodies, and means for circulating at least one of a medium and a sample between the two porous bodies. 9. The culture apparatus according to claim 1, wherein the case has means for pushing out the contents contained in the porous body or the porous body itself out of the case, or sucking the contents from outside the case. 10. The culture apparatus according to claim 1, wherein a drug having a bactericidal action is held in the case, and a means for killing microorganisms after culture by contacting the drug with the porous body is provided. .