JPH09154566A - Tool for testing food poisoning - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a tool capable of detecting and identifying food poisoning bacteria according to simple operations by arranging a microbial collecting part having a specific structure in a test tubelike hollow container. SOLUTION: This tool for testing food poisoning is obtained by using a lid unit 2 of a hollow cylindrical container 1 having an opening at one end as a microbial collecting part and the tip of a rodlike member (6a) as a microbial collecting end (6b) and arranging a baglike member 4 enclosing a culture medium 3 suitable for culturing food poisoning bacteria in the upper part of the lid unit 2. A pH indicator for detecting a change in pH due to the microbial propagation is contained in the culture medium 3, which is dropped into the interior of the container by breaking the baglike member 4.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a food poisoning bacterium detection instrument capable of easily and easily detecting and / or identifying food poisoning bacteria, and more specifically, detecting / identifying food poisoning bacteria by a simple operation. The present invention relates to an apparatus for detecting food poisoning bacteria, which is particularly suitable for personal and home use.

[0002]

2. Description of the Related Art Ingestion of food and drink, that is, "food" motion, is
It is indispensable for animals including humans. Despite recent advances in science and sophistication of technology,
Even now, it is difficult to completely prevent "food poisoning" which is an addiction caused by ingestion of food and drink. On the other hand, the prevention of such food poisoning has become more important due to the recent enforcement of the Product Liability Law (PL Law).

Based on the cause, food poisoning is caused by natural poisons (fugu poison, mushroom poison, etc.), food poisoning by decay of food and drink (decomposed products of food and drink), and bacteria (staphylococcus, salmonella) to food and drink. , Vibrio parahaemolyticus) or food poisoning due to breeding.

Among these food poisoning, food poisoning due to natural poison can be easily avoided by avoiding ingestion of food or drink containing the causative poison. Further, since it is generally easy to detect spoilage of food and drink based on changes in the appearance and smell of the food and drink, it is often easy to avoid food poisoning due to spoilage of food and drink. In contrast to these two types of food poisoning, bacterial food poisoning (ie bacterial food poisoning)
In the above, the contamination or reproduction of the bacterium in food and drink is usually invisible, and the detection of the contamination or reproduction is extremely difficult without the involvement of a specialist. Therefore,
This bacterial food poisoning is the most difficult food poisoning to avoid.

The occurrence of food poisoning in a facility that supplies a large amount of food and drink, such as a cafeteria, a bento catering business, and a lunch center,
Symptoms such as severe vomiting, diarrhea, abdominal pain, and high fever occur in a large number of people, so that particularly serious problems (such as long-term work suspension of the facility) are likely to occur.

[0006]

Conventionally, a means for preventing food poisoning has been a general preventive measure (for example, keeping kitchen utensils such as kitchen knives and mana plates clean, and keeping fingers and hands of people involved in cooking clean. Keep, etc.).

[0007] Even in the present situation, when food poisoning occurs, the causative bacterium is detected and confirmed by an investigation (cultivation of bacteria adhering to food and drink considered to be the cause, etc.) at a health center having jurisdiction over the place of occurrence, that is, after the fact. I had no choice but to take an effective measure. In addition, detection of causative bacteria based on the survey of public health centers
It was customary to require a significantly long time (for example, about 1 to 2 weeks) for confirmation.

[0008] Furthermore, for the investigation of public health centers, "prescribed procedures"
Not only was it underutilized, it was not always underutilized, as it was not only required to take the money, but also because of problems with honor and commercial rights (for example, the complete loss of the customer due to "reputation").

An object of the present invention is to provide a food poisoning bacteria detection instrument capable of detecting and identifying food poisoning bacteria by a simple operation.

Another object of the present invention is to provide a food poisoning bacteria detection instrument suitable for personal and home use.

Still another object of the present invention is to provide a food poisoning bacterium detection instrument that makes it extremely easy to take measures beforehand or voluntarily.

[0012]

Means for Solving the Problems As a result of earnest research, the present inventor has arranged a medium and a fungal collection part which are to be provided with culture conditions suitable for culturing a specific food poisoning bacterium when they are in contact with each other, And, at the time of culture for detecting food poisoning bacteria,
It has been found that it is extremely effective to achieve the above-mentioned object to construct a detection instrument in which these specific culture medium and bacteria collection section are provided so as to be in contact with each other.

The food poisoning bacterium detection instrument of the present invention is based on the above findings, and more specifically, a hollow container having an opening at one end; a bacterium sampling portion arranged in the hollow container; It is characterized in that it comprises at least a medium which is arranged so as to come into contact with the bacterium collecting portion during culturing and which has selectivity for culturing food poisoning bacteria.

In the above-described food poisoning bacterium detection instrument of the present invention, when not in use, the culture medium and the bacterium sampling section which should provide culture conditions suitable for culturing food poisoning bacteria are provided in non-contact with each other.

On the other hand, at the time of culturing for detecting food poisoning bacteria, these specific medium and the bacteria collecting portion are brought into contact with each other to give a state suitable for the propagation of food poisoning bacteria. In the present invention, the medium is further put into a state suitable for breeding food poisoning bacteria of a specific species (for example, Staphylococcus aureus), and other food poisoning bacteria (for example, Salmonella) is substantially not grown. To be done.

Therefore, a suitable bacterial culture means (for example,
The use of a simple incubator) and bacterial growth detection means (for example, a pH indicator that detects a pH change due to bacterial growth) facilitates detection of food poisoning bacteria. When a detection instrument of a mode that provides culture conditions having selectivity for a food poisoning bacterium of a specific species (for example, Staphylococcus aureus) is further used, detection / identification of a food poisoning bacterium of a specific species (type of food poisoning bacterium) Identification) is easy.

In the present invention, "selectivity" in culturing a bacterium
Preferably has a culture selectivity for at least one or more kinds of bacteria selected from the group consisting of "Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella" that accounts for almost all (about 99%) of bacteria causing food poisoning. However, the distinction between these three types of bacteria may not be essential.
That is, when the presence of at least one of the three types of bacteria is confirmed, it is possible to substantially remove all of the three types of bacteria by adopting a predetermined disinfecting means described later. Because it is possible.

When the food poisoning bacteria can be detected and identified as described above (for example, spontaneously), a means suitable for removing the food poisoning bacteria (for example, cooking utensils and the like, trade name "HIBIDEN" ( 0.2-0.5% chlorohexidine solution) or a disinfection method of cleaning with water after wiping with an inverted soap)
By adopting, it becomes possible to easily and effectively prevent food poisoning.

The bacterium detection device of the present invention, if necessary,
It is configured to confer culture selectivity for Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella, which are the "three major" food poisoning bacteria (ca. 99% of bacterial food poisoning is due to either of these).

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will now be described in detail with reference to the drawings as necessary.

FIG. 1 shows a schematic side sectional view of a preferred embodiment of the food poisoning bacterium detection instrument of the present invention.

With reference to FIG. 1, the food poisoning bacteria detecting device in this embodiment comprises a hollow cylindrical container having an opening at its upper end, and a lid 2 which can be removably fitted in the opening. To be done.

At the upper part of the lid 2 (opposite to the container opening of the lid 2), a bag shape containing a medium 3 suitable for culturing a specific food poisoning bacterium (for example, Staphylococcus aureus) therein. The member 4 is arranged. The medium 3 further contains a pH indicator (for example, phenol red) that detects a pH change due to bacterial growth.

On the other hand, a perforated plate 5 having one or more (preferably a plurality of) holes is arranged on the lower side of the bag-shaped member 4 (opening side of the container), and the bag-shaped member 4 serves as a container. It prevents it from falling to the opening side.

Further, at a substantially central portion on the bottom surface side of the lid body 2, there is provided a bacteria collecting portion 6 extending substantially perpendicular to the opening surface of the container. The bacteria collecting section 6 is composed of a rod-shaped member 6a provided on the side closer to the lid 2 and a bacteria-collecting end 6b arranged at the tip of the rod-shaped member (the side farther from the lid 2). If necessary, the bacteria collection end 6b has an antibiotic (for example, Aztreonam) having a property of effectively suppressing the growth of bacteria other than the above-mentioned specific food poisoning bacteria (for example, Staphylococcus aureus). A combination of polymyxin B and fluconazole) is given.

In the present invention, it is also possible to dispose an antibiotic separately from the above-mentioned container, the medium 3 or the bacteria collecting section 6. In the embodiment in which the antibiotics are separately arranged in this way, for example, a disc-shaped member (not shown; preferably made of a "porous body") to which the antibiotics are applied is provided inside the container 1 as necessary. You may arrange.

When such a food poisoning bacteria detecting instrument of the embodiment shown in FIG. 1 is used, as shown in FIG. 2A, the lid 2 having the bacteria collecting portion 6 is taken out from the container. Then, FIG.
As shown in (b), the bacteria collection end 6b at the tip of the bacteria collection unit 6
Is rubbed against an inspection target (for example, a cutting board) 10 to collect the bacteria present in the inspection target 10 at the bacteria collection end 6b.

After inserting the lid 2 having the bacteria collecting portion 6 into the container again, as shown in FIG. 2 (b), the bag-like member 4 (for example, using a pressing device not shown, the lid 2 The bag-shaped member 4 by squeezing it from the outside of the bag)
The medium 3 contained in the container is dropped inside the container. As a result, the bacteria collecting end 6b of the bacteria collecting portion 6 is immersed in the medium 3. At this time, the materials other than the culture medium, such as the fragments of the bag-shaped member 4, are prevented from falling by the perforated plate 5 and do not fall inside the container.

As described above, since the bacteria collecting end 6b is provided with an antibiotic having a specific selectivity, the bacteria collecting end 6b of the bacteria collecting part 6 is thus immersed in the medium 3. Then, certain food poisoning bacteria (eg Staphylococcus aureus)
The reproduction of bacteria other than is substantially suppressed. Therefore, when the bacterial detection instrument in such a state is subjected to culture by an appropriate bacterial culture means (for example, a simple incubator), specific food poisoning bacteria (for example, Staphylococcus aureus) are selectively propagated. It will be. The culture conditions at this time are not particularly limited, but usually, the conditions of 37 ° C. and about 16 to 24 hours are sufficient.

Presence or absence of discoloration of the pH indicator contained in the medium 3 (and the degree of discoloration) after the above-mentioned culture operation.
Based on the above, it is possible to easily detect and identify a specific food poisoning bacterium (for example, Staphylococcus aureus) (identify the type of food poisoning bacterium).

Next, the structure of each part of the food poisoning bacteria detection instrument of the present invention will be described.

(Container) The container is preferably made of a transparent or translucent material. More specifically, the container is made of an inorganic material such as glass or plastic (for example, polystyrene, polyethylene terephthalate (PET),
The container can be formed using an organic material such as an acrylic resin or a polyolefin resin), but considering the weight and the possibility of damage during centrifugation or the like, the container is preferably made of plastic.

The shape of the container is not particularly limited as long as it has an opening at the upper end and the lid 2 can be removably attached to the opening. More specifically, for example, in addition to the illustrated test tube, it may be in a container shape such as a flask shape or a bottle shape.

(Lid) The material forming the lid 2 is not particularly limited, but from the viewpoint of reducing the possibility of breakage of the container and enhancing the adhesion with the container, the lid 2 is better than the container. It is preferably composed of a soft material. More specifically, when the container is made of a hard plastic or a hard resin such as polystyrene or PET, the lid 2 is made of a soft plastic or a soft resin such as polyolefin (used to include an elastic body or an elastomer).
It is preferably composed of

(Medium) In the present invention, the type and composition of the medium 3 are not particularly limited as long as desired food poisoning bacteria can be cultivated, and the combination with an antibiotic, a pH indicator, etc. described later is suitable. It can be appropriately selected in consideration of the properties (reactivity, compatibility, etc.).

In the present invention, examples of the medium for Staphylococcus aureus include mannitol salt (modified) medium, Baird-
Parker medium, Tellurite-Glycine medium, Phenylethanol-
Azide medium, chocolate agar medium, blood agar medium, heart infusion agar medium, etc. can be used,
From the viewpoint of compatibility with an antibiotic for selection of Staphylococcus aureus described later, it is preferable to use a mannitol salt (modified) medium.

On the other hand, examples of the medium for Vibrio parahaemolyticus include salt polymyxin (modified) medium and Cellobiose-Polym.
ixin-Colistin medium, Thiosulfate-Citrate-Bile Salts
-Sucrose medium, TCBS agar medium, MacConkey agar medium, blood agar medium, etc. can be used, but salt polymyxin (modified) medium is used in view of compatibility with antibiotics for selection of Vibrio parahaemolyticus described below. It is preferable.

Further, examples of the Salmonella medium include xylose / lysine (modified) medium, Mannitol-Lys
ine-Crystal Violet-Brilliant medium, Salmonella-Shige
lla medium, Deoxychocolate-Citrate-Lactose-Sucrose medium, DHL agar medium, MacConkey agar medium, etc. can be used, but xylose / lysine (modified ) It is preferable to use a medium.

In the present invention (by a gel-like medium or the like)
Although it is possible to detect and identify food poisoning bacteria by using food poisoning bacteria cultured in a colony, it is easy to detect and identify food poisoning bacteria based on color change due to pH change etc. in general cafeterias and homes. From this point of view, it is preferable to use a liquid medium that gives a uniform culture system. The medium 3 may have a certain degree of viscosity as long as it does not substantially prevent the detection / identification of food poisoning bacteria based on color change or the like.

(Antibiotic) In the present invention, it is possible to impart a certain degree of food poisoning bacteria selectivity to the above-mentioned medium 3 itself, but from the standpoint of ensuring this selectivity, it is necessary. Therefore, it is preferable to use a combination of antibiotics that effectively suppress the growth of bacteria other than the specific food poisoning bacteria to be detected and confirmed.

As described above, "selectivity" in culturing bacteria
Preferably has a culture selectivity for one or more kinds of bacteria selected from the group consisting of "Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella", which accounts for almost all (about 99%) of bacteria causing food poisoning. However, discrimination between these three types of bacteria is not essential. However, from the point that it is possible to take detailed measures according to the characteristics of each of these food poisoning bacteria, it is possible to have "selectivity of culture" that can distinguish between "Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella". preferable.

In the present invention, any antibiotic can be used without particular limitation as long as it is an antibiotic which can effectively suppress the growth of bacteria other than the target specific (one or more) food poisoning bacteria. For example, the following antibiotics are preferably used. In the following, the concentrations shown together with the respective antibiotics are suitable concentrations at the time of culturing food poisoning bacteria (when integrated with the medium).

<A Staphylococcus aureus-selective antibiotic> Aztreonam 1 to 15 μg / ml Polymyxin B 1 to 15 μg / ml Fluconazole 1 to 10 μg / ml A combination of the above three kinds of antibiotics can be preferably used. .

<Vibrio parahaemolyticus-selective antibiotic> Polymyxin B 1 to 15 μg / ml Fluconazole 1 to 10 μg / ml Potassium tellurite 1
20 μg / ml A combination of the above three kinds of antibiotics can be preferably used.

<Salmonella Selective Antibiotic> Fluconazole 1 to 10 μg / ml (position of application of antibiotic) In the present invention, as long as the above antibiotic functions together with the medium 3 during the culture of food poisoning bacteria, the antibiotic The application position is not particularly limited.

More specifically, for example, the antibiotic is the medium 3
It may be added in advance, or may be coated on the inner wall of the container, and is applied to a predetermined position of the bacteria collecting portion 6 (position of the bacteria collecting portion 6 that can come into contact with the medium 3 during culture). May be. Two or more of these application positions may be combined as needed. It goes without saying that in these aspects, the above-mentioned disc-shaped member can be omitted.

When the antibiotic is added to the medium 3 in advance, the inactivation of the antibiotic in the medium is taken into consideration (for example, at about 30 to 35% at room temperature for 1 year). It is preferable to add a large amount of antibiotics to cover the decrease in the activity of the antibiotics.

On the other hand, in a mode in which an antibiotic is applied to a predetermined position of the bacteria collecting section 6, the rod-shaped portion 6a of the bacteria collecting section 6 is used.
It is preferable that it is provided (at a position where it can come into contact with the medium 3 at the time of culturing) and / or at the bacteria collection end 6b at the tip of the bacteria collection part 6. When the antibiotic is applied to the bacteria collecting end 6b, the bacteria collecting end 6b itself may be impregnated, or the bacteria collecting end 6b may be composed of an inner layer (rod-like member 6a side) and an outer layer (for collecting bacteria). An antibiotic may be applied to the inner layer portion as a multi-layered structure composed of two or more layers.

(Bag-shaped member) In the present invention, the bag-shaped member 4 can stably and surely enclose the medium 3 during storage and transportation of the food poisoning bacteria detection instrument, and the medium 3 can be used during the culture of food poisoning bacteria.
The material, shape, etc. of the bag-shaped member 4 are not particularly limited as long as the predetermined amount can be reliably discharged.

More specifically, the material of the bag-shaped member 4 may be a hard material such as glass or hard plastic, or a soft material such as soft plastic such as polyethylene or polypropylene or a flexible material such as paper. Although it may be present, it is preferably made of a transparent or translucent material from the viewpoint of easily confirming the presence of the medium to be encapsulated and the color tone thereof.

In the present invention, during the culture of food poisoning bacteria,
The means for releasing the medium 3 from the bag-shaped member 4 is not particularly limited, as long as the medium 3 is arranged at a position suitable for the culture.
More specifically, for example, when releasing the culture medium 3 from the bag-shaped member 4, any means such as punching, cutting, and breaking the bag-shaped member 4 can be used. When the bag-shaped member 4 is punched or cut, a member such as a needle-shaped member or a blade-shaped member (neither is shown) that facilitates punching or cutting is provided on the inner wall of the lid body 2 as necessary. You may arrange. On the other hand, in the mode in which the culture medium 3 is released by breaking the bag-shaped member 4, the bag-shaped member 4 is preferably made of a hard material (glass, hard plastic, etc.) from the viewpoint of facilitating the breaking.

(Bar-shaped member) In the present invention, the bar-shaped member 6
a is stably holding the bacteria collection end 6b having a function of directly collecting food poisoning bacteria while avoiding contamination,
In addition, it is a member having a so-called auxiliary function of holding the bacteria collecting end 6b at a position suitable for contacting with the medium 3 during culture of bacteria. The material, length, shape and the like of the rod-shaped member 6a are not particularly limited as long as the functions of stable holding and contact position securing are substantially achieved. The rod-shaped member 6a is preferably made of a flexible plastic in terms of prevention of contamination and flexibility in collecting bacteria.

(Bacteria collection end) In the present invention, the bacteria collection end 6
b is a member having a function of directly collecting food poisoning bacteria and contacting the medium 3 during culture of the bacteria to provide conditions suitable for culturing the food poisoning bacteria. The material, length, shape and the like of the bacteria collection end 6b are not particularly limited, as long as such a collection function and a contact function are substantially achieved. More specifically, for example, the bacteria-collecting end 6b may be a mere tip of the rod-shaped member 6a. However, from the viewpoint of easily achieving the above-mentioned collecting function / contact function, it is possible to collect the bacteria. The end 6b preferably has a surface with an increased surface area. Examples of such a "surface with increased surface area" include a surface (for example, an "earpick" or a tip similar to a spatula) provided with one or more recesses (cuts, uneven surfaces, etc.), or a porous surface. The surface etc. are mentioned.

In the embodiment in which the bacteria collecting end 6b (at least the surface portion thereof) is made of a porous material, a xerogel-like porous body or a fibrous body is preferably used. As the fibrous body, for example, filter paper (paper), cotton (absorbent cotton, etc.), punch felt, woven fabric, knitted fabric, non-woven fabric and the like can be used without particular limitation.

(Disc Member) In the present invention, the disc member, which is arranged as necessary, holds an antibiotic, and is brought into contact with the medium 3 at the time of culturing the bacterium so that the condition suitable for culturing a specific type of food poisoning bacterium is set. It is a member having the function of providing. The material, length, shape and the like of the disk member are not particularly limited as long as the holding function and the contact function are substantially achieved. More specifically, for example, the disc member may be a simple plate or a planar member, but in terms of easily achieving the above-mentioned holding function / contact function, the disc member has a large surface area. It is preferred to have an increased surface. Examples of such a “surface having an increased surface area” include a surface provided with one or more recesses (cuts, uneven surfaces, etc.), a porous surface, or the like.

In the embodiment in which the disk member (at least the surface portion thereof) is made of a porous material, a xerogel-like porous body or a fibrous body is preferably used. As the fibrous body, for example, filter paper (paper), cotton (absorbent cotton, etc.), punch felt, woven fabric, knitted fabric, non-woven fabric and the like can be used without particular limitation.

(Additives) Various additives can be added to the above-mentioned medium 3 as required. In the embodiment of the present invention in which the detection of food poisoning bacteria is confirmed by the pH based on the growth of the bacteria, it is preferable to add a pH indicator to the medium in advance. When such a pH indicator is used, acidic substances (lactic acid, etc.) are produced by the growth of bacteria and the pH of the medium changes, and the color tone of the pH indicator changes based on this, making it easy to confirm the reproduction. Becomes

The pH indicator usable in the present invention is not particularly limited. From the viewpoint of pH change range, color tone, etc., for example, phenol red, cresol red, bromthymol blue, bromcresol purple, triphenyl tetrazolium, blue tetrazolium and the like can be preferably used. These pH indicators can be used alone or in combination of two or more as needed. Furthermore, by using pH indicators each having a different color tone (when unchanged) for the medium for detecting different bacterial species, it becomes easy to recognize the bacterial species to be detected at a glance by these color tones. Therefore, it is preferable.

(Aseptic condition) The food poisoning bacteria detection instrument of the present invention as described above is preferably substantially aseptic until just before use (at least inside thereof) from the viewpoint of accuracy of inspection. When the device of the present invention is manufactured into a sterile state after production, known physical means (for example, sterilization by irradiation with electromagnetic waves such as ultraviolet rays and γ rays) and / or chemical means (for example, ethylene oxide and the like). Gas sterilization) can be used without particular limitation.

After the above sterilization, the food poisoning bacteria detection instrument of the present invention is preferably stored in a bag (not shown) made of a film or the like. At this time, food poisoning bacteria detection device (Fig. 1)
May be stored integrally in the bag, or if necessary, the container and the lid 2 (+ bacteria collecting section 6) may be stored in separate bags. .

FIG. 3 shows an example of another embodiment of the food poisoning bacteria detection instrument of the present invention. In the embodiment shown in FIG. 3, when the culture medium 3 falls into the container 1, the culture medium 3 propagates along the rod-shaped member 6a.
A “guide member” 20 having a function of guiding the falling of the perforated plate is provided below the perforated plate 5. The configuration of FIG. 3 is the same as the configuration of FIG. 1 except that the guide member 20 is provided.

The material, shape, size, etc. of the guide member 20 are not particularly limited as long as the guide member 20 has a guide function of promoting the dropping of the medium 3 propagating along the rod-shaped member 6a. It is preferable that at least a part of the guide member 20 has an inner diameter smaller than the inner diameter of the container 1 from the viewpoint of facilitating the drop of the culture medium 3 propagating along the rod-shaped member 6a.

More specifically, for example, the guide member 20
As shown in FIG. 3, it is preferable that the lower end is made of a "vane" -like member whose lower end is slightly inclined toward the center of the container 1 (that is, the lower end has an inner diameter smaller than the inner diameter of the container 1). Such “tilt” can be omitted.

Hereinafter, the present invention will be described more specifically with reference to Examples.

[0065]

Example 1 (Production of Staphylococcus aureus detection instrument) Referring to FIG. 1, the following materials were used to produce a food poisoning bacteria detection instrument having the following dimensions.

Container 1: polyethylene (thickness 0.6 mm), inner diameter 8.
5 mm, height 150 mm bottomed hollow cylinder Lid 2: polyethylene (thickness 0.6 mm), inner diameter 10
mm, 53 mm long hollow cylinder with a bottom Bag-shaped member 4: made of glass (thickness 0.5 mm), inner diameter 7.5
mm hollow cylinder with a length of 40 mm Perforated plate 5: polyethylene (thickness 1 mm), diameter 6 m
m disk-shaped body rod-shaped body 6a: made of polyethylene, diameter 2 mm, length 135
mm Microbial collection end 6b: absorbent cotton, 5 mm diameter, 15 mm long "cotton swab" tip disk member: 1 mm thick, 6 mm diameter filter paper (when the antibiotics shown in the following (Table 1) are integrated with the medium) Aztreonam 10 μg / ml, polymyxin B 8 μg / ml, fluconazole 5 μg / ml
A disc member (not shown) disposed between the bottom of the container 1 and the bacteria collection end 6b was impregnated with the amount of the concentration of ml. ) Medium 3: Medium having the composition (excluding antibiotics) shown below (Table 1)

[0067]

[Table 1]

After the detection instrument for Staphylococcus aureus having the above-mentioned constitution was manufactured, it was sterilized by irradiating with γ ray at 1 to 30 ° C. for 1 minute. The sterilized instrument was stored in a bag made of a polyethylene film (thickness 100 μm) vapor-deposited with aluminum for light shielding until just before use.

Example 2 (Preparation of Vibrio parahaemolyticus detection instrument) With reference to FIG. 1, the procedure of Example 1 was repeated except that the composition of the medium 3 and the antibiotics were changed as shown in the following (Table 2). A bacterium detection instrument for Vibrio parahaemolyticus was prepared, sterilized, and stored. On this occasion,
When the antibiotics shown in (Table 2) were integrated with the medium, polymyxin B 10 μg / ml, fluconazole 5 μg
/ Ml, and potassium tellurite at a concentration of 5 μg / ml, the disk member was impregnated.

[0070]

[Table 2]

Example 3 (Preparation of Salmonella Detection Instrument) Referring to FIG. 1, medium 3
A detection instrument for Salmonella was prepared, sterilized, and stored in the same manner as in Example 1 except that the composition and the antibiotics were changed as shown below (Table 3). At this time, the disc member was impregnated with the antibiotics shown in (Table 3) in an amount such that the concentration of fluconazole was 5 μg / ml when integrated with the medium.

[0072]

[Table 3]

Example 4 (Confirmation of Selectivity of Staphylococcus aureus Detection Medium) Example 1
The selectivity of the Staphylococcus aureus detection medium used in 1. was confirmed as follows.

To 1.5 ml of the Staphylococcus aureus detection medium 3 used in Example 1, 22 kinds of bacteria shown in Table 4 below were adjusted to a concentration of about 10 ⁶ cells / ml at the beginning of culture. Dispersed in the air for 24 hours at 37 ° C for 4 hours.
The presence or absence of proliferation of various bacteria was judged by the presence or absence of discoloration of the pH indicator phenol red (contained in the above medium) after culturing for 8 hours and 72 hours. When the color tone of the culture medium after the culture was “yellow”, it was defined as “positive +” (with bacterial growth), and when the color tone of the medium was “red”, it was defined as “negative-” (without bacterial growth).

[0075]

[Table 4]

The results obtained are shown in Table 4. As is clear from the results in Table 4, when the Staphylococcus aureus detection medium of this example was used, the growth of Gram-negative bacteria and fungi was caused by the antibiotics and antifungals adsorbed on the medium 3 and the disk 7. It was confirmed that it was effectively suppressed by the agent.

Of the bacteria tested above, S. epidermi
Since dis did not decompose mannitol contained in the medium 3, the color tone of the medium did not change.

According to the experiments of the present inventor, other than the Staphylococcus aureus, Coagulas are the only bacterial species that are positive in this medium.
e negative Staphylococcus (CNS), E.faecalis,
Although E.faecium was found to exist, these bacteria could be distinguished from Staphylococcus aureus by the phosphatase or coagulase test (literature name: hygiene inspection guideline, staphylococcal test guideline).

In the phosphatase test, a phosphatase detection disk (filter paper on which 4-methylumbelliferyl phosphate was adsorbed) was used to incubate in the above medium, and then fluorescence of 4-methylumbelliferone was irradiated with ultraviolet rays ( It was possible to confirm the existence by irradiating the same.

Example 5 (Confirmation of Selectivity of Vibrio parahaemolyticus Detection Medium) The selectivity of the Vibrio parahaemolyticus detection medium used in Example 2 was confirmed as follows.

To 1.5 ml of the Vibrio parahaemolyticus detection medium 3 used in Example 2, 22 kinds of bacteria shown in Table 5 below were added so that the concentration of the bacteria was about 10 ⁶ cells / ml at the beginning of the culture. Disperse in air at 37 ° C for 24 hours, 48 hours each
Propagation of various bacteria was judged by the presence or absence of discoloration of the pH indicators bromthymol blue and cresol red (containing the above medium) after culturing for 72 hours and 72 hours. When the color tone of the culture medium after the culture was "yellow", it was defined as "positive +" (with bacterial growth), and when the color tone of the medium was "green", it was defined as "negative-" (without bacterial growth).

[0082]

[Table 5]

The results obtained are shown in Table 5. As is clear from the results in Table 5, when the culture medium for detecting Vibrio parahaemolyticus of this example was used, the growth of Gram-positive bacteria was suppressed by the antibiotics, and most of the Gram-negative bacteria and fungi did not grow. , Antibiotics adsorbed on disk 7 (polymyxin B, fluconazole, and potassium tellurite)
Was confirmed to be effectively suppressed by.

According to the experiments conducted by the present inventor, other than the Vibrio parahaemolyticus, V. cholera was found to be a positive bacterial species in this medium.
e, V. vulnificus, Preteus spp, etc. were found, but these bacteria were tested for oxidase (cytochrome oxidase reaction, literature name: Kovacs, Nature (Londo
n), 178 , 703, 1956), it was possible to distinguish from Vibrio parahaemolyticus.

Example 6 (Confirmation of Selectivity of Salmonella Detection Medium) The selectivity of the Salmonella detection medium used in Example 3 was confirmed as follows.

Salmonella detection medium 3 used in Example 3
22 kinds of bacteria shown in Table 6 below were dispersed in 1.5 ml of the above so as to have a concentration of about 10 ⁶ cells / ml at the beginning of the culture, and at 37 ° C. in the air for 24 hours and 48 hours, respectively. The presence or absence of breeding of various bacteria was judged by the presence or absence of discoloration of the pH indicator bromocresol purple (containing the above medium) after culturing for 72 hours. When the color tone of the culture medium after the above culture is "black", it is defined as "positive +" (there is the proliferation of bacteria), and the color tone of the medium is "purple or yellow".
The case was designated as "negative-" (no bacterial reproduction). Hydrogen sulfide-producing bacteria such as Salmonella turned the medium black by changing the thiosulfate contained in the medium with ammonium iron citrate.

[0087]

[Table 6]

The obtained results are shown in Table 6. As is clear from the results in Table 6, when the Salmonella detection medium of this example was used, the growth of Gram-positive bacteria was suppressed by the antibiotic, and the growth of the fungus was the antibiotic adsorbed on the disk 7. It was confirmed that it was effectively suppressed by the substance (fluconazole).

Experiments conducted by the present inventor have revealed that some Gram-negative bacteria change the color tone of this medium from purple to yellow. An example of such a change in color tone is shown below.

* Change to black: Salmonella * Change to yellow: Escherichia coli, Citrobac
ter spp, Klebsiella spp, Enterobacter spp, Proteu
s spp * Changes to purple: Shigalla spp, Serratia sp
p, Pseudomonas spp Example 7 (Use of Staphylococcus aureus detection instrument) After breaking the bag of the detection instrument for Staphylococcus aureus obtained in Example 1 and taking out the instrument, the lid 2 (and the bacteria collection part 6) Take out the container from the container (Fig. 2 (a)), and attach the bacteria collection end 6b to a cooking utensil (such as a cutting board).
The surface was rubbed several times (FIG. 2 (b)).

Then, the lid 2 is inserted into the container again and the bacteria collecting end 6b is held at a predetermined position inside the container.
Using a compression device (a wide clip-shaped "dividing device") not shown, the bag-shaped member 4 is strongly pressed from the outside of the lid body 2 to be destroyed, and the medium 3 contained in the bag-shaped member 4 is stored in a container. Dropped inside. Thereby, the bacteria collection end 6b of the bacteria collection unit 6
Is immersed in the medium 3 (at this time, materials other than the medium such as the fragments of the bag-shaped member 4 are prevented from falling into the container by the perforated plate 5).

When the bacteria-collecting end 6b of the bacteria-collecting section 6 was immersed in the medium 3 in this way and incubated at 37 ° C. for 16 to 24 hours, the color tone of the medium 3 was changed by a certain cooking tool (cutting board). It changed from red to yellow. That is, the presence of Staphylococcus aureus was confirmed.

The cookware in which the presence of Staphylococcus aureus was confirmed was wiped several times with gauze dipped in an aqueous solution of 0.2 to 0.5% chlorohexidine (trade name "HIBIDEN", manufactured by Sumitomo Pharmaceutical Co., Ltd.). After that, it was washed with tap water for 10 minutes.

In this way, the Staphylococcus aureus detection test was conducted on the cooking utensils (cutting board) after being wiped with the inverted soap and washed with water in the same manner as above, but the color tone of the medium 3 remained yellow and did not change. No Staphylococcus aureus was detected. That is, it was confirmed that Staphylococcus aureus was effectively removed by the above disinfection treatment.

Example 8 (Use of Vibrio parahaemolyticus and Salmonella detection instrument) In the same manner as in Example 7, except that the Vibrio parahaemolyticus detection instrument and the Salmonella detection instrument obtained in Examples 2 and 3 were used, When the presence of food poisoning bacteria was confirmed, the presence of Vibrio parahaemolyticus or Salmonella was confirmed for a certain cooking utensil.

When these cooking utensils were wiped with reverse soap and washed with water in the same manner as in Example 7, the presence of Vibrio parahaemolyticus or Salmonella was not confirmed after the disinfection treatment. That is, it was confirmed that the above-mentioned disinfection treatment effectively removes Vibrio parahaemolyticus or Salmonella.

[0097]

As described above, according to the present invention, there is provided a food poisoning bacteria detecting instrument capable of detecting and identifying food poisoning bacteria by a simple operation.

Since the food poisoning bacteria detection instrument of the present invention is extremely easy to use for personal and home use, voluntary detection of food poisoning bacteria without the involvement of experts such as public health centers, and preventive measures against food poisoning based on the detection. Is possible.

[Brief description of the drawings]

FIG. 1 is a schematic side cross-sectional view showing one embodiment of a food poisoning bacteria detection instrument of the present invention.

FIG. 2 is a schematic diagram showing an example of a method of using the food poisoning bacteria detection instrument of FIG.

FIG. 3 is a schematic side sectional view showing another embodiment of the food poisoning bacteria detection device of the present invention.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 ... Container main body, 2 ... Lid body, 3 ... Medium, 4 ... Bag-shaped member, 5
... Perforated plate, 6 ... Bacteria collecting part, 6a ... Rod-shaped member, 6b ... Bacteria collecting end, 20 ... Guide member.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Reference number within the agency FI Technical display location // G01N 33/48 G01N 33/48 S (72) Inventor Naomi Osawa Onta Town, Higashimurayama City, Tokyo 5-38-15 Celtec Laboratory Co., Ltd.

Claims (13)

[Claims] 1. A hollow container having an opening at one end, a bacterium collection section arranged in the hollow container, and a bacterium collection section arranged so as to come into contact with the bacterium collection section during culture of the bacterium, and food poisoning. A food poisoning bacteria detection instrument comprising at least a medium having selectivity for culturing bacteria. 2. The food poisoning bacterium detection device according to claim 1, further comprising an antibiotic so that at least a part of the bacterium is in contact with the medium during culture of the bacterium. 3. The food poisoning bacteria detection instrument according to claim 2, wherein the antibiotic is placed in the hollow container. 4. The food poisoning bacteria detection instrument according to claim 2, wherein the antibiotic is added to the medium. 5. The food poisoning bacteria detecting device according to claim 2, wherein the antibiotic is applied to the bacteria collecting portion. 6. The food poisoning bacteria detection instrument according to claim 3, wherein the inner wall of the hollow container is coated with the antibiotic. 7. The food poisoning bacteria detection instrument according to claim 3, wherein the antibiotic is applied to another member arranged in the hollow container. 8. The device for detecting food poisoning bacteria according to claim 3, wherein the bacteria collecting portion has a multiple structure including at least an inner material and an outer material, and the antibiotic is arranged on the inner material. 9. The medium comprises a substance that changes a color tone based on a change caused by the growth of bacteria.
The food poisoning bacteria detection instrument described. 10. The food poisoning bacteria detecting device according to claim 1, which has a culture selectivity for at least one kind of bacteria selected from the group consisting of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella. 11. The culture medium is contained in a bag-shaped member, and the bag-shaped member is arranged in the lid body.
The food poisoning bacteria detection instrument described. 12. The food poisoning bacteria detection instrument according to claim 11, wherein a perforated member is arranged between the bag-shaped member and the hollow container. 13. The food poisoning bacteria detection instrument according to claim 12, further comprising a guide member disposed below the perforated member, the guide member accelerating the dropping of the medium propagating through the bacteria collecting portion into the hollow container.