JP3519793B2 - Microorganism testing equipment - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microbial testing instrument used for examining the presence of microorganisms in a sample.

[0002]

2. Description of the Related Art Blood is originally in a sterile state when it is in the body. However, when an infection occurs, bacteria may enter the blood and further cause sepsis. If sepsis is suspected, it is necessary to immediately check for the presence of bacteria. On the other hand, when transfusing a patient with a weakened immune system, such as a terminal cancer patient, it is important to check that the blood to be transfused is sterile before use.

[0003] Conventionally, when performing a microbial test for sterility of a sample such as blood, a sample is taken from a container containing the sample with a syringe or the like in a clean bench and used for a microbial test containing a medium. Transfer to a device,
After culturing at 40 ° C., turbidity, hemolysis, change in color of reagent due to gas, etc. are used as indicators for determination.

The problem here is that, when it is determined that microorganisms are present, there is always a possibility that the sample itself is contaminated with microorganisms and that microorganisms are contaminated during the operation of the microorganism inspection. That is. There is a possibility that various bacteria may be mixed in during the operation of the microorganism test.When the needle connected to the container containing the sample is pierced into the rubber stopper of the container, the other bacteria not originally contained in the sample due to contact with fingers or the like. May be mixed in the microbiological test equipment. This is
Disinfecting the rubber stopper of the sample container and the rubber stopper of the microbiological test instrument with alcohol or the like does not solve the problem.

Further, when the sample is blood, it is unavoidable that blood traces or splashes on the surface of the rubber stopper of the container. This may be touched by a person engaged in the inspection, and there is a risk of being infected with hepatitis virus, AIDS virus, or the like.

[0006]

SUMMARY OF THE INVENTION The present invention has been made to solve the problems of contamination of microorganisms in a microbiological test instrument and splashing of a sample in a microbiological test, and the sample is aseptically and without splashing the sample. An object of the present invention is to provide a device for microbiological examination, which can collect water.

[0007]

As a result of repeated studies to solve the above problems, it was found that the above-mentioned problems can be achieved by the following microbiological test instruments, and the present invention was completed. It was

(1) A bottomed tube capable of accommodating a sample, a tube made of a thermoplastic resin whose base end communicates with the opening of the bottomed tube, and a rupture which interrupts the communication between the bottomed tube and the tube. A microbe inspection instrument comprising a possible sealing member, wherein the bottomed tube has a culture medium, and the tube has a filter that allows gas to pass but does not pass microorganisms at a tip thereof. Equipment.

(2) A bottomed tube capable of accommodating a sample, a thermoplastic resin tube communicating with the opening of the bottomed tube, and a rupturable seal for blocking the communication between the bottomed tube and the tube. A device for microbiological examination comprising a stop member, wherein the bottomed tube has a medium, a filter through which a gas passes but a liquid does not pass, and a pH indicator, and the pH indicator is removed from the medium by the filter. A device for microbiological examination characterized by being arranged so as to be isolated.

(3) The above-mentioned tube has a filter through which gas passes but does not pass through microorganisms at its tip, and its proximal end can communicate with the opening of the bottomed tube. .

(4) The microbiological test instrument according to any one of (1) to (3), wherein the pressure inside the bottomed tube is reduced.

(5) The microorganism testing instrument according to any one of (1) to (4), wherein the material of the tube is polyvinyl chloride resin.

(6) The microorganism testing instrument according to any one of (1) to (5), wherein the material of the bottomed tube is either glass or hard plastic.

[0014]

EXAMPLES Aseptic according to the present invention refers to a state of being sterilized by heat, high-pressure steam or the like, or a state of being in an aseptic state not exposed to the outside air. Normal operation in the clean bench is not included because it is exposed to the outside air.

The microbiological test instrument of the present invention will be described below in detail with reference to the preferred embodiments shown in the accompanying drawings.

FIG. 1 and FIG. 2 show an embodiment of the microbiological test instrument of the present invention. FIG. 1 is a microbiological test instrument 1A.
FIG. FIG. 2 is a vertical cross-sectional view of the microorganism testing instrument 1A. A rubber plug 30 is fitted into the opening 21 of the bottomed tube 20 of the microbe inspection instrument 1A, and a crowned member 80 is crowned from the outer surface of the rubber plug 30 to the outer peripheral surface of the opening 21. A base end 52 of a tube 50 is connected to the rubber stopper 30 via a sealing member 40. Sealing member 40
Can be broken by a finger or the like, and when broken, the lumen of the tube 50 and the lumen of the bottomed tube 20 communicate with each other. At the tip 51 of the tube 50, a first filter 60 that is permeable to gas but impermeable to microorganisms is attached. Also, the bottomed tube 2
The 0 lumen has medium 70 and is depressurized. Furthermore, due to the high-pressure steam sterilization, the inside of the microorganism testing instrument 1A is in a sterile state.

The shape of the bottomed tube 20 is not particularly limited.
The material is glass or a hard plastic that can be sterilized by autoclave, and one having good internal visibility is preferably used. As the hard plastic, a material having low air permeability is preferable from the viewpoint of airtightness, and for example, acrylonitrile-based thermoplastic resin, polyvinylidene chloride-polyvinyl chloride copolymer, polyethylene terephthalate, hard vinyl chloride and the like can be used. , Polyvinylidene chloride-polyvinyl chloride copolymers or materials based on them are preferred. The size of the bottomed tube 20 depends on the amount of medium,
It is determined in consideration of the amount of the sample to be collected and the volume in the tube and is not particularly limited, but may be, for example, about 30 to 80 ml. The degree of decompression inside the bottomed tube 20 is not particularly limited, but is determined in consideration of the amount of sample to be collected and the volume in the tube.

The rubber stopper 30 is fitted into the bottomed tube 20, and further, a crowned member 80 is crowned from the outer surface of the rubber stopper 30 to the outer peripheral surface of the opening 21 to hermetically seal the inner cavity of the bottomed tube 20. It is increasing the nature. The lower end of the sealing member 40 in FIG. 2 is connected to the rubber plug 30, and the inner cavity of the lower end of the sealing member 40 and the inside of the bottomed tube 20 communicate with each other. As the material of the rubber plug 30, a so-called cross-linked rubber such as isoprene rubber (IR), butyl rubber (IIR), silicone rubber (Q), or a thermoplastic elastomer can be used.

The crown member 80 is not essential, but it is preferable to provide it in order to maintain the airtightness of the bottomed tube 20. The crowned member 80 may be crowned so as to maintain the airtightness of the bottomed tube 20, and the material is not particularly limited, but aluminum, aluminum alloy, or the like is preferable.
Instead of the crowned member 80, the bottomed tube 20 and the rubber plug 3
0 may be adhered to each other with an adhesive or the like.

FIG. 3 is an enlarged vertical sectional view showing a structural example of the sealing member 40. The sealing member 40 is composed of a short tube 41 made of a flexible material to be described later, and a tubular body 42 fitted in the short tube 41 and closed at one end by a solid columnar portion 43. There is. Short tube 4
One end of the tube 50 is connected to the upper end of FIG. The lower end of the cylindrical body 42 in FIG. 3 is connected to the rubber stopper 30. A thin and fragile breaking portion 44 is formed on the outer periphery of the cylindrical body 42. The solid columnar portion 4 together with the short tube 41 from the outside of the short tube 41 by fingers or the like.
By bending 3 to break the breaking portion 44 and separating the solid columnar portion 43, the internal flow path of the sealing member 40 is opened. As the material of the cylindrical body 42, for example, hard polyvinyl chloride, hard plastic such as polycarbonate, or the like can be used. The material of the short tube 41 may be the same as the material shown in the tube 50 described later.

The tube 50 preferably has the same diameter as the tube communicating with the opening of the container in which the sample is stored. Particularly, a plurality of bags for storing and storing blood components are connected by the tube. It is preferable that the tube has the same diameter as the tube used in the bag connecting body. The inner diameter of the tube used for the bag connecting body is 2.0 to
It is about 4.5 mm. Although details will be described later, it is preferable to have such a size because it is easy to aseptically connect to a tube connected to a container in which the sample is stored when collecting the sample.

The material of the tube 50 is preferably a thermoplastic resin, preferably polyvinyl chloride or a material mainly containing it. If the tube 50 is made of polyvinyl chloride,
It is preferable because sufficient flexibility and flexibility can be obtained, easy handling, and good connectivity. The type and content of the plasticizer used in each tube are not particularly limited.

The first filter 60 is attached to the tip 51 of the tube 50 and allows gas to pass but does not allow microorganisms to pass.
With this configuration, since the tip is not closed, the lumen of the tube 50 is not crushed and fused by high-pressure steam sterilization, and microorganisms do not pass through. Therefore, the lumen of the tube 50 is aseptic. Can be maintained in a state.
The first filter 60 can be manufactured from a sintered body obtained by molding polyethylene, polypropylene, fluororesin, or the like into powder and then heat-compressing and molding. In addition,
Instead of the sintered body, a membrane filter in which a polyester resin aggregate is coated with polyvinyl chloride may be used. As the material of the membrane filter, other materials such as polytetrafluoroethylene laminated on a polyethylene non-woven fabric, or an acetate film coated with a silicone resin can be used. The pore size of the first filter 60 is preferably 0.45 μm or less, more preferably 0.22 μm or less.

The medium 70 may be appropriately selected depending on the microorganism to be detected. The amount of the medium 70 is not particularly limited, but may be about 15 to 40 ml, for example. For the purpose of detecting aerobic bacteria, brain heart infusion broth, trypticase soy broth, columbia broth, trypto soy broth, ordinary broth, heart infusion broth, thioglycolic acid medium and the like can be used. For the purpose of detecting anaerobic bacteria,
Thioglycolic Acid Medium, Thiogel Medium, Columbia Broth, Cooked Sheet Medium, TEP (Triple-Extract-Pep
tone) Broth medium etc. can be used. If necessary, a bactericidal inhibitor such as sodium polyanetholesulfonate (SPS), an anticoagulant such as heparin, a reagent for detecting microorganisms such as a pH indicator, etc. may be added to the medium 70.

FIG. 4 is a vertical cross-sectional view showing another embodiment of the microorganism testing instrument of the present invention. Microbial inspection instrument 1
The plan view of B is the same as that of the microbe inspection instrument 1A of FIG. The microbe inspection instrument 1B has the same configuration as that of the microbe inspection instrument 1A, but the pH of the inner wall surface of the bottomed tube 20 is
A difference is that there is an indicator 90 and a second filter 100, which is permeable to gas but impermeable to liquid, isolates the pH indicator 90 from the medium 70.

The pH indicator 90 may be applied to the inner wall surface of the bottomed tube 20 or the second filter 100, but it is more preferable to use a support such as filter paper which is impregnated with the pH indicator or combined with the support. Easy to distinguish and preferable. As the pH indicator 90, a pH indicator and a fluorescent pH indicator that can be visually discriminated can be used, but they may be appropriately selected because they differ depending on the pH of the gas to be detected. For example, when detecting carbon dioxide, bromthymol blue, phenol red or the like can be used as the pH indicator 90.

The second filter 100 is permeable to gas but impermeable to liquid, covers one side of the pH indicator 90, and has a bottomed tube 2
Liquid-tightly attached to the inner wall surface of 0, and the pH indicator 90 is added to the medium 7
Separated from zero. As the material, a non-woven fabric made of ES fibers, that is, polypropylene for the inner core portion and polyethylene for the outer layer portion, or a laminate of non-woven fabric made of polypropylene such as Tyvek (trade name) can be used. With such a configuration, the pH indicator 90 is not dimmed by the medium 70 or the sample, so that the determination can be reliably performed.

Next, a method for collecting a sample using the microorganism testing instrument 1A of the present invention will be described. FIG. 5 is a soft plastic bag 120 containing a sample. A base end 152 of a tube 150 made of the same material as the microorganism testing device 1A of the present invention is connected to the container 110 of the soft plastic bag 120 and communicates with the inside thereof, and a tip 151 of the tube 150 is closed.

Microbial test instrument 1A or 1B of the present invention
As a method of aseptically connecting the tube 50 of No. 3 and the tube 150 of the soft plastic bag 120 containing the sample, the following heating connection means 200 described in Japanese Patent Publication No. 3-68819 is used. There is a method. FIG. 6 is a perspective view schematically showing a configuration example of the heating connection means 200, and FIGS. 7 to 10 are perspective views showing a connecting step between the tube 50 and the tube 150 using the heating connection means 200. As shown in these figures, the heating connection means 200
The wafer 20 between the pair of holders 201 and 202.
3 is a structure in which the tube 50 of the device 1A or 1B for microbiological examination and the tube 150 of the container 120 in which the sample is housed are stacked between the holders 201 and 202 in the opposite direction, and the wafer is heated. These are cut and melted at 203, one holder 201 is moved, and the tube 50 and the tube 1 are removed except for the wafer 203.
50 and 50 are fused together.

The heating connection means 200 will be described in detail below with reference to FIGS. As shown in FIG. 7, the tube 50 and the tube 150 are piled up in parallel for a certain length so that the tubes 50 and 150 are oriented in opposite directions, and the two holders 20
1 and 202 in the grooves 205 and 206, and the holder pieces 211, 212 and 221, 222 are closed, and the tube 50 and the tube 150 are attached to the tube holding portions 207 and 2
Hold and fix at 08.

Next, as shown in FIG.
Is heated to a temperature equal to or higher than the melting temperature of the tube 50 and the tube 150, the tube 50 and the tube 150 supported in parallel by the blade 225 thereof are cut, and the cut ends of the tube 50 and the tube 150 are cut by the heat of the wafer 203. To melt. At this time, since the cut ends of the tube 50 and the tube 150 are in a melted or softened state, the heat keeps the aseptic state.

Next, as shown in FIG. 9, while maintaining the molten state of the cut ends of the tubes 50 and 150, one holder 201 is moved in the arrangement direction of the tubes 50, and the cut tubes 50 and Tube 150
Stop and fix at the position where the cut ends face each other.

Next, as shown in FIG.
3 is pulled out in a direction perpendicular to the tubes 50 and 150, and one holder 201 is pressed toward the other holder 202. Thereby, the cut ends of the melted tube 50 and the tube 150 are bonded (fused) to each other. Even when the tube is connected, the wafer 203 is sterilized by heat, so that the aseptic state is maintained. In addition, the tubes to be connected are not limited to having the same composition, and may be compatible with each other.

By the above-mentioned method, a microbe inspection instrument 1A
Alternatively, after the 1B tube 50 and the tube 150 of the soft plastic bag 120 containing the sample are connected, the breaking portion 44 of the sealing member 40 is broken by a finger or the like. Then, since the inner diameter of the bottomed tube 20 of the microorganism testing instrument 1A or 1B is reduced, the sample flows into the bottomed tube 20. When the required amount of sample is obtained, the tube 50 is closed near the sealing member 40 by heat fusion or the like, and the center of the closed portion is cut to separate the soft plastic bag 120. By cutting in this way, the end of the cut tube remains closed, the sterility is maintained and the sample does not leak.

The microbiological test instrument 1A or 1B from which the sample is collected is cultured at an appropriate temperature. Although the temperature varies depending on the microorganism, it is usually preferably 37 to 40 ° C. The number of culture days varies depending on the type of microorganism and the detection method, but is usually about 1 to 10 days.

The presence or absence of microorganisms is determined by using turbidity of the medium, hemolysis, change in color of pH indicator due to gas generation, appearance of bacterial mass, and the like. The determination based on the generation of gas such as carbon dioxide gas includes a method using a change in color of the pH indicator as an index and a method using an infrared spectroscope. In the microorganism testing instrument 1B, the pH indicator 9 can be discriminated by the above index.
The change in color of 0 can be judged visually or by a measuring device, and particularly when a fluorescent pH indicator is used, the sensitivity is high, so that it can be judged in a shorter period of time.

The microbiological test instrument of the present invention comprises blood, urine,
A liquid such as a peritoneal dialysate collected in a bag can be aseptically collected from each bag, and a microbial test such as sterility can be performed.

[0038]

EFFECTS OF THE INVENTION Since a tube is connected to the opening of the bottomed tube of the microorganism testing instrument of the present invention, by aseptically connecting to the tube connected to the container in which the sample is stored, Samples can be collected aseptically without contamination by external bacteria and without splashing. Then, by culturing this microbe inspection instrument, it is possible to surely perform the microbe inspection without making an erroneous determination due to contamination of various bacteria. Further, since the inside of the bottomed tube is depressurized, when the sealing member is broken, the sample automatically flows into the bottomed tube. Furthermore, since the tube is made of polyvinyl chloride, it has good flexibility and flexibility, and it is easy to connect with the tube of the bag in which the liquid is collected. Microbial tests can be performed.

Further, in the microbiological test instrument of the present invention, the filter that allows gas but not liquid separates the pH indicator from the medium. For this reason, the pH indicator is not dimmed in color by the medium or sample, so that the determination can be reliably performed.

[Brief description of drawings]

FIG. 1 is a front view showing an embodiment of a microorganism testing instrument of the present invention.

FIG. 2 is a vertical cross-sectional view showing an embodiment of the microorganism testing instrument of the present invention.

FIG. 3 is a vertical cross-sectional view of a sealing member.

FIG. 4 is a front view showing another embodiment of the microorganism testing instrument of the present invention.

FIG. 5 is a front view of a soft plastic bag containing a sample.

FIG. 6 is a perspective view schematically showing a configuration example of heating connection means for connecting a tube.

FIG. 7 is a perspective view showing a tube connecting step using the heating connecting means of FIG.

8 is a perspective view showing a tube connecting process using the heating connecting means of FIG.

9 is a perspective view showing a tube connecting step using the heating connecting means of FIG. 6. FIG.

FIG. 10 is a perspective view showing a tube connecting process using the heating connecting means of FIG.

[Explanation of symbols]

1A, B Microbiology test equipment 20 bottomed tube 21 opening 30 rubber stopper 40 sealing member 41 short tube 42 cylinder 43 Solid Column Section 44 fracture 50 tubes 51 Tube tip 52 proximal end of tube 60 First Filter 70 medium 80 Crowned member 90 reagents 100 second filter 110 containers 120 Soft plastic bag 150 tubes 151 tube tip 152 Tube End 200 Heating connection means 201 holder 202 holder 203 wafer 205 groove 206 groove 207 tube holder 208 tube holder 211 Holder piece 212 Holder piece 221 Holder piece 222 Holder piece 225 blade

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Claims (4)

(57) [Claims] 1. A bottomed tube capable of accommodating a sample, a thermoplastic resin tube whose base end communicates with an opening of the bottomed tube, and a breakable tube which blocks the communication between the bottomed tube and the tube. For detecting microorganisms, comprising a sealing member and a culture medium in the bottomed tube, wherein the tube has a filter that allows gas to pass but does not pass microorganisms at the tip. Equipment. 2. A bottomed tube capable of accommodating a sample, a thermoplastic resin tube communicating with the opening of the bottomed tube, and a rupturable seal for blocking communication between the bottomed tube and the tube. A device for microbiological examination comprising a member, wherein the bottomed tube has a medium, a filter through which a gas passes but a liquid does not pass, and a pH indicator, and the pH indicator is isolated from the medium by the filter. A device for microbiological testing, which is characterized by being arranged. 3. The microorganism testing instrument according to claim 2, wherein the tube has a filter through which gas passes but does not pass through microorganisms, and a proximal end of the tube can communicate with the opening of the bottomed tube. 4. The pressure inside the bottomed tube is reduced.
The microbiological test instrument according to any one of 1 to 3.