JP2010057390A - Test implement - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a test implement capable of easily preparing a selectively bleeding medium for campylobacter in a short time, with no need of preparing any sterilizing vessel for preparing the medium. <P>SOLUTION: The test implement is such that, a sealable packaging bag 10 formed of a transparent flexible resin material is packed with such an amount of base medium powder 12 as to be necessary for a single test, a first internal packaging bag 14 encapsulated with such an amount of horse laked blood as to be necessary for a single test and burstable by externally applied force, and a second internal packaging bag 16 encapsulated with such an amount of a supplement liquid as to be necessary for a single test and burstable by externally applied force, and the resultant packaging bag 10 is sealed up. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a Campylobacter test tool used in preparing a enrichment medium for selective enrichment culture of Campylobacter in the test of Campylobacter that detects Campylobacter that is one of the causative bacteria of food poisoning.

The test | inspection which detects Campylobacter in foodstuffs, such as meat, is implemented by performing each operation in order of sample preparation, enrichment culture, isolation culture, and identification as follows, for example.
First, a sample of 25 g (or 25 mL) is collected from a specimen, and the sample is placed in a stomat bag, and then 100 mL of the enrichment medium is added and homogenized (homogenized) to prepare the sample. As the enrichment medium, a Preston enrichment medium is usually used (for example, see Patent Document 1), and a Bolton enrichment medium is used when there is a high possibility of germ damage (for example, see Patent Document 2). .) Next, when a Preston enrichment medium is used, enrichment culture is performed at a temperature of 42 ± 1 ° C. for 24-48 hours under microaerobic conditions (or aerobic conditions). In addition, when the Bolton enrichment medium is used, after incubation for 4 hours at a temperature of 35 ± 1 ° C. under microaerobic conditions (or aerobic conditions), increase for 24 to 44 hours at a temperature of 42 ± 1 ° C. Perform bacterial culture or enrichment culture at a temperature of 42 ± 1 ° C. for 24-48 hours. Next, the enriched culture is streaked on a separation medium and separated and cultured at a temperature of 42 ± 1 ° C. for 24-48 hours. As the separation medium, for example, mCCDA medium is used. Bacteria are identified after this separation culture. For example, the colonies on the medium are observed, some of the suspicious colonies are collected, smeared on a blood agar medium and an equivalent non-selective medium with low sugar content, and a temperature of 42 ± 1 ° C. -Incubate for 48 hours. Then, identification and identification are performed by confirming the bacterial form by Gram staining, the presence or absence of catalase positive, the presence or absence of oxidase positive, the presence or absence of positive in latex agglutination test, and the like.

The enrichment medium used in the above Campylobacter test is prepared, for example, as follows. That is, the Preston enrichment medium is composed of a basal medium powder prepared by suspending basal medium powder containing peptone, Lab-Lemco powder and sodium chloride in purified water, dissolving by heating, horse lysate, and acetone / sterilization. Prepared by aseptically mixing Campylobacter selective supplement and Campylobacter growth supplement dissolved in purified water (1: 1) and dispensing into sterile containers. The Bolton enrichment medium is composed of a basal medium sterilized by suspending a basal medium powder containing meat peptone, lactalbumin hydrolyzate, yeast extract, sodium chloride, etc. Prepared by aseptically mixing blood with a Bolton selective supplement dissolved in ethyl alcohol / sterilized purified water (1: 1) and dispensed into sterile containers.
JP 2005-516627 A (pages 7-8, 18) JP 2005-511014 A (page 7)

  At business establishments such as clinical laboratories, food factories, and medical institutions that conduct Campylobacter examinations, basal medium powders, growth supplements, and selected supplements have been purchased and stored in the past. The enrichment medium was prepared by a simple operation, and enrichment culture was performed using the prepared enrichment medium. Therefore, weigh out the amount of basal medium powder necessary for one test, sterilize the basal medium, dissolve selective supplements in a solvent, etc., and select basal medium and horse hemolyzed blood in the container It was necessary to prepare the enrichment medium by stirring and mixing with supplements, etc., and these operations took time and effort. Moreover, there is a problem that it is necessary to prepare a sterilization container.

  The present invention has been made in view of the circumstances as described above. A selective enrichment medium for Campylobacter can be prepared easily and in a short time, and a sterile container is particularly prepared for the preparation of the enrichment medium. An object of the present invention is to provide a Campylobacter inspection tool which does not need to be performed.

  The invention according to claim 1 is a Campylobacter test tool for preparing an enrichment medium for testing for detecting Campylobacter, and is a sealable package formed of a transparent or translucent resin material having flexibility or flexibility. Inside the body, the amount of basal medium powder necessary for one test is accommodated, and the amount of horse hemolyzed blood necessary for one test and the amount of supplement solution required for one test are enclosed and packaged. An inner package that is ruptured by a pressing force applied from the outside of the body is accommodated, and the package is sealed, and when used, the package is opened and sterilized purified water is injected into the package, Dissolve the internal basal medium powder with sterilized purified water, rupture the internal package, mix the basal medium, horse lysate and supplement solution, and select and enrich Campylobacter in the package. They were prepared, characterized by enrichment culture by injecting a specimen to a packaging body to which the enrichment medium housed.

  The invention according to claim 2 is the inspection tool according to claim 1, wherein the inner package includes a first inner package in which horse hemolyzed blood is enclosed, and a second inner package in which supplement liquid is enclosed. It is divided into two internal packaging bodies, and the first internal packaging body and the second internal packaging body are each ruptured during use.

  The invention according to claim 3 is the test tool according to claim 1 or 2, wherein the basal medium powder is a Preston basal medium, and the supplement liquid is Campylobacter growth supplement and Campylobacter selective supplement, To do.

  The invention according to claim 4 is the inspection tool according to claim 1, wherein the basal medium powder is a Preston basal medium, the supplement liquid is Campylobacter growth supplement and Campylobacter selective supplement, and the inner package is Three internal packages, a first internal package enclosing horse hemolyzed blood, a second internal package encapsulating Campylobacter growth supplement, and a third internal package encapsulating Campylobacter selective supplement The first inner package, the second inner package, and the third inner package are each ruptured during use.

  The invention according to claim 5 is the inspection tool according to claim 1 or 2, wherein the basal medium powder is a Bolton basal medium, and the supplement liquid is a Campylobacter selective supplement.

  The invention according to claim 6 is the inspection tool according to any one of claims 1 to 5, wherein the package containing the basal medium powder and the inner package is sealed and used after being frozen. Sometimes, the horse hemolyzed blood and supplement liquid enclosed in the inner package are respectively thawed.

  According to a seventh aspect of the present invention, in the inspection tool according to the sixth aspect, γ-rays, X-rays, or electron beams are irradiated to the package that contains the basic medium powder and the inner package and is sealed and frozen. The basal medium powder is sterilized.

  The invention according to claim 8 is the inspection tool according to any one of claims 1 to 7, wherein the inner package is formed of a film material in a sealed bag shape and at least a part of the peripheral edge is weakly bonded. It is characterized by that.

In the inspection tool of the invention according to claim 1, the package is opened, sterilized purified water is injected into the package, and the basal medium powder in the package is dissolved with the sterilized purified water. Then, by pressing the outer surface of the package with a finger and applying a pressing force to the inner package in the package, the inner package is ruptured, and horse hemolyzed blood and supplement liquid flow out from the inner package. Then, the selective enrichment medium of Campylobacter is prepared in an amount necessary for one test in the package by mixing the basal medium, the horse lysate and the supplement solution and mixing them sufficiently. Campylobacter enrichment culture is performed by injecting the specimen into the package containing the enrichment medium, sealing the package, and homogenizing.
Therefore, the use of the test tool of the invention according to claim 1 makes it possible to easily prepare a selective enrichment medium for Campylobacter in a short time, and it is also necessary to prepare a sterilized container especially for the preparation of the enrichment medium. No.

  In the inspection tool of the invention according to claim 2, the first inner package is formed by pressing the outer surface of the package with a finger and applying a pressing force to the first inner package and the second inner package in the package, respectively. The body and the second inner package are ruptured, the horse hemolyzed blood flows out from the first inner package, and the supplement liquid flows out from the second inner package.

  In the inspection tool of the invention according to claim 3, the Preston enrichment medium can be prepared easily and in a short time.

  In the inspection tool of the invention according to claim 4, the outer surface of the package is pushed with a finger to apply a pressing force to the first inner package, the second inner package and the third inner package in the package. Accordingly, the first inner package, the second inner package, and the third inner package are each ruptured, and horse hemolyzed blood flows out from the first inner package, and Campylobacter is discharged from the second inner package. The growth supplement flows out and the Campylobacter selective supplement flows out from the third inner package. And with this test | inspection tool, a Preston enrichment culture medium can be prepared easily and in a short time.

  In the inspection tool of the invention according to claim 5, the Bolton enrichment medium can be prepared easily and in a short time.

  In the inspection tool of the invention according to claim 6, it is possible to stably store the equine hemolyzed blood and the supplement solution stored in the inner package for a long period of time.

  In the inspection tool of the invention according to claim 7, when it is difficult to aseptically enclose the basal medium powder in the package, the basal medium powder is sterilized after encapsulating in the package. Can do.

  In the inspection tool of the invention according to claim 8, by pressing the outer surface of the package with a finger and applying a pressing force to the inner package in the package, the peripheral edge of the inner package is easily peeled off. Burst.

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the best embodiment of the present invention will be described with reference to the drawings.
FIG. 1 is a plan view showing an example of an embodiment of the present invention, with a part of an inspection tool broken.

  This inspection tool is composed of a basal medium powder 12 and two inner packaging bags in a packaging bag 10 formed of a transparent (or translucent) resin material having flexibility (or flexibility), for example, nylon, polyethylene or the like. 14 and 16 and the packaging bag 10 is hermetically sealed. In this example, the packaging bag 10 is a stand bag that can stand on its own. In the vicinity of the upper side of the packaging bag 10, a fastener 18 is inserted between two film-like sheets constituting the packaging bag 10, and a linear male member and a linear female member constituting the fastener 18 are in a film form. Each is welded to the sheet. And the packaging bag 10 is sealed and opened by engaging and releasing the linear male member and the linear female member of the fastener 18. Two film-like sheets constituting the packaging bag 10 are thermally bonded to each other on the upper side of the packaging bag 10, and the packaging bag 10 is sealed by the thermal bonding unit 20. Further, a tear line 22 is printed between the thermal bonding portion 20 and the fastener 18. Instead of printing the cut line 22, notches that are slightly cut in a V shape from both sides of the packaging bag 10 may be formed between the thermal bonding portion 20 and the fastener 18. As shown in the plan view of FIG. 2, the sealed packaging bag 10 is obtained by cutting the film-like sheet with scissors or the like along the cut line 22 or tearing the two film-like sheets at the notch forming portion. Opened.

  The amount of the basal medium powder 12 enclosed in the packaging bag 10 is an amount necessary for one inspection. Further, the amount of horse hemolyzed blood necessary for one inspection is enclosed in the first inner packaging bag 14, and the amount of supplement necessary for one inspection is contained in the second inner packaging bag 16. Liquid is enclosed. Each of the first inner packaging bag 14 and the second inner packaging bag 16 applies a pressing force to the first inner packaging bag 14 and the second inner packaging bag 16 by pressing the outer surface of the packaging bag 10 with a finger. It is formed so as to burst. For example, each of the first inner packaging bag 14 and the second inner packaging bag 16 is formed in a sealed bag shape with a film material, and a part of the peripheral edge portions 24 and 26 is weakly bonded by thermal bonding or the like. have. Then, when a force is applied to the first inner packaging bag 14 and the second inner packaging bag 16, the weakly bonded peripheral edges 24 and 26 are easily peeled off, and the first inner packaging bag 14 and Each of the second inner packaging bags 16 is ruptured, horse hemolyzed blood flows out of the first inner packaging bag 14, and supplement liquid flows out of the second inner packaging bag 16.

  The composition of the basal medium powder includes peptone, Lab-Lemco powder and sodium chloride for the preparation of Preston enrichment medium, and meat peptone, lactalbumin hydrolyzate, yeast extract, sodium chloride for the preparation of Bolton enrichment medium. Etc. The supplement solution enclosed in the second inner packaging bag 16 is Campylobacter growth supplement (composition: sodium pyruvate, sodium metabisulfite and iron sulfate (hydrated salt)) and Preston for the preparation of Preston enrichment medium. It is prepared by dissolving Campylobacter selective supplements (composition: polymyxin B, rifampicin, trimethoprim and cycloheximide) with acetone / sterilized purified water (1: 1). For the preparation of a Bolton enrichment medium, Bolton selection supplements (composition: cefoperazone, vancomycin, trimethoprim and cycloheximide) are dissolved in ethyl alcohol / sterilized purified water (1: 1).

  The packaging bag 10 in which the basal medium powder 12, the first inner packaging bag 14 and the second inner packaging bag 16 are accommodated and sealed is subjected to a freezing treatment, and the horse hemolyzed blood in the first inner packaging bag 14 and The supplement liquid in the second inner packaging bag 16 is frozen. Although the equine hemolyzed blood and supplement solution cannot be stored for a long period of time at refrigeration or at room temperature, it is possible to stably store the test tool for a long period of time by freezing the packaging bag 10. This inspection tool is frozen and stored in clinical laboratories, food factories, medical institutions, and other establishments, and is taken out when selective enrichment culture of Campylobacter is required, and enclosed in the first inner packaging bag 14. The horse hemolyzed blood and the supplement solution enclosed in the second inner packaging bag 16 are used after being thawed. Although the operation of sealing the basal medium powder 12 into the packaging bag 10 is performed aseptically, if this is difficult, the basal medium powder 12 and the first inner packaging bag are contained inside the packaging bag 10. 14 and the second inner packaging bag 16 are sealed and the packaging bag 10 is frozen, and then the basal medium powder 12 is sterilized by irradiating the packaging bag 10 with γ-rays, X-rays or electron beams. At this time, the basal medium powder can be sterilized without reducing the performance of the medium by appropriately adjusting the dose of γ rays and the like.

  As a more specific preparation example of this test tool, when preparing a 100 mL Bolton selective enrichment medium, a sterilized Bolton selective enrichment bouillon (powder) is added to a sterilized transparent stand bag with fasteners. Add 76g. In a sterile sachet, 5 mL of sterile equine hemolyzed blood is placed and sealed. In another sterile sachet, 2 mg of cefoperazone, 2 mg of vancomycin, 2 mg of trimethoprim and 5.1 mg of cycloheximide are added with 1 mL of ethyl alcohol / sterilized purified water (1: 1). Seal the dissolved sterile solution. And after putting a sterile small bag in a stand bag, the opening part of a stand bag is heat-bonded and sealed.

  When inspecting Campylobacter using the above-described inspection tool, after thawing frozen horse hemolyzed blood and supplement solution to return to room temperature, first, along the cut line 22 near the upper side of the packaging bag 10 The film-like sheet is cut, the packaging bag 10 is opened, and sterilized purified water is injected into the packaging bag 10 so that the concentration becomes 1 times. After the upper side of the packaging bag 10 is sealed with the fastener 18, the basal medium powder 12 inside the packaging bag 10 is dissolved with sterile purified water. Next, by pressing the outer surface of the packaging bag 10 with a finger and applying a pressing force to the first inner packaging bag 14 and the second inner packaging bag 16 in the packaging bag 10 respectively, the first inner packaging bag 14 and Each of the second inner packaging bags 16 is ruptured so that the horse hemolyzed blood flows out from the first inner packaging bag 14 and the supplement solution flows out from the second inner packaging bag 16. Then, the selective enrichment medium of Campylobacter is prepared in an amount necessary for one test in the packaging bag 10 by mixing the basal medium, the horse hemolyzed blood and the supplement solution and mixing them sufficiently. Then, an amount of specimen necessary for one test is injected into the packaging bag 10 containing the enrichment medium, and lightly homogenized as usual so that as much air as possible does not remain in the packaging bag 10. The packaging bag 10 is sealed with a fastener 18. After this, enrichment culture of Campylobacter is performed in the packaging bag 10 as usual.

  In the above-described embodiment, the horse hemolyzed blood is sealed in the first inner packaging bag 14 and the supplement liquid is sealed in the second inner packaging bag 16, but in one inner packaging bag. A supplement solution may be enclosed together with the horse hemolyzed blood, and one of the inner packaging bags may be accommodated in the packaging bag 10. For example, for the preparation of Preston enrichment medium, three internal packaging bags are used, horse hemolyzed blood is enclosed in the first internal packaging bag, Campylobacter growth supplement is enclosed in the second internal packaging bag, The Preston Campylobacter selective supplement may be enclosed in the third inner packaging bag. Moreover, when accommodating a some internal packaging bag in the inside of the packaging bag 10, it is not necessary to burst several internal packaging bags simultaneously. For example, in the enrichment culture using the Bolton enrichment medium, after the basic medium powder 12 in the packaging bag 10 is dissolved with sterilized purified water, first, only the first inner packaging bag 14 in the packaging bag 10 is pressed. Is added to rupture only the first inner packaging bag 14 and cause the horse hemolyzed blood to flow out from the first inner packaging bag 14. In this state, after culturing at a temperature of 35 ± 1 ° C. for 4 hours under microaerobic conditions, the second inner packaging bag 16 is ruptured by applying a pressing force to the second inner packaging bag 16. Then, the solution of the Bolton selective supplement that is an antibiotic is allowed to flow out of the second inner packaging bag 16, and the supplement solution is thoroughly mixed with the medium. In this state, the temperature is 42 ± 1 ° C. for 24-44 hours. Enrichment culture may be performed.

It is a top view which shows one example of embodiment of this invention and shows the state which fractured | ruptured some inspection tools. It is a top view which shows the state which opened the packaging bag of the test | inspection tool shown in FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Packaging bag 12 Basal medium powder 14 1st inner packaging bag 16 2nd inner packaging bag 18 Fastener 20 Thermal bonding part 22 Cut line 24, 26 The weakly adhere | attached peripheral part of an internal packaging bag

Claims (8)

An amount of basal medium powder necessary for one test is contained in a sealable package formed of a transparent or translucent resin material having flexibility or flexibility, and for one test. It contains an internal package that contains the required amount of horse hemolyzed blood and the amount of supplement liquid required for one test, and is ruptured by the pressing force applied from the outside of the package, and the package is sealed. ,
At the time of use, the packaging body is opened, sterilized purified water is injected into the packaging body, the basal medium powder in the packaging body is dissolved with sterilized purified water, the inner packaging body is ruptured, and the basal medium and horse hemolyzed blood And a supplement solution, a selective enrichment medium of Campylobacter is prepared in the package, a specimen is injected into the package containing the enrichment medium, and the enrichment culture is performed. The inner package is divided into two inner packages, a first inner package enclosing horse hemolyzed blood and a second inner package enclosing a supplement liquid,
The Campylobacter inspection tool according to claim 1, wherein the first inner package and the second inner package are each ruptured during use. The Campylobacter test tool according to claim 1 or 2, wherein the basal medium powder is a Preston basal medium, and the supplement solution is Campylobacter growth supplement and Campylobacter selective supplement. The basal medium powder is a Preston basal medium, and the supplement liquid is Campylobacter growth supplement and Campylobacter selective supplement,
The inner package includes a first inner package enclosing horse hemolyzed blood, a second inner package enclosing Campylobacter growth supplement, and a third inner package enclosing Campylobacter selective supplement Are divided into three inner packagings,
The Campylobacter inspection tool according to claim 1, wherein the first inner package, the second inner package, and the third inner package are each ruptured during use. The Campylobacter examination tool according to claim 1 or 2, wherein the basal medium powder is a Bolton basal medium, and the supplement liquid is a Campylobacter selective supplement. The packaged body containing the basal medium powder and the inner package and sealed is frozen and stored, and the equine hemolyzed blood and the supplement solution enclosed in the inner package are thawed when used. The Campylobacter inspection tool according to claim 5. The Campylobacter test according to claim 6, wherein the basal medium powder is sterilized by irradiating the basal medium powder and the inner package containing the sealed and frozen package with γ-ray, X-ray or electron beam. Tools. The Campylobacter inspection tool according to any one of claims 1 to 7, wherein the inner package is formed of a film material in a sealed bag shape and at least a part of a peripheral edge thereof is weakly bonded.

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