JP6576489B2 - Method for collecting microorganisms and genes, and method for extracting the same - Google Patents

Description

  The present invention relates to a wipe-type inspection performed in the food field, the medical field, and the like, and relates to a method for collecting microorganisms and genes, and a method for extracting the same.

  Conventionally, in the food field, a wiping inspection of bacteria and viruses (hereinafter sometimes simply referred to as “microorganisms”) has been performed in a work space from the viewpoint of safety and health. In the inspection, microorganisms present on the surface of the inspection object, including cooking utensils in the work space (for example, cutting boards, kitchen knives, etc.) and other equipment such as refrigerators, sinks, shelves, door knobs, handrails, floors, etc. Further, a wiping inspection method is carried out in which a swab or the like impregnated with a predetermined amount of a diluent such as phosphate buffer or physiological saline is wiped off, and the wiped microorganisms and genes are detected by a technique such as culture. Also in the medical field, a wiping inspection method is carried out in which the surface of an object to be inspected such as a medical device or a medical facility is wiped and the wiped microorganisms and genes are detected by a technique such as culture.

  As a wiping device for microorganisms and genetic testing (hereinafter sometimes simply referred to as “microbiological testing”), a tube equipped with a microorganism wiping carrier that holds an aqueous solution inside the lid is used, and the microorganisms after wiping by centrifugation are used. Has been disclosed (for example, see Patent Document 1 below).

Japanese Patent Laid-Open No. 2006-85083

  Conventionally, when testing using a method such as culturing of microorganisms or genes (eg, gene amplification method (polymerase chain reaction method (PCR method))), for pathogenic viruses and pathogens with low abundance A method of concentrating the collected liquid after the recovery is employed. For example, in the conventional method, the amount of the diluted solution is set to a few mL to 10 mL unit, but a multi-stage concentration operation is required to collect a small amount of microorganisms from such a diluted solution. However, when a multi-stage concentration operation is required, the work becomes complicated, and there are some problems such as work mistakes, recovery loss, and recovery variations. On the other hand, in the technique disclosed in Patent Document 1, it has been studied to solve the problem, but the amount of wiping liquid is described as about 0.5 mL to 1 mL, and examination is performed by a gene amplification method or the like. Therefore, it is assumed that a concentration step is still necessary. In this respect, in order to further improve the accuracy of the test result, it is desirable that the concentration process is as few as possible or the concentration process is not performed.

  Furthermore, it is preferable that the operation of collecting microorganisms or the like can be easily performed without using complicated means and apparatuses. For example, it is desirable that a worker can easily collect microorganisms and the like without requiring special knowledge in a store or factory that handles food. The surface to be inspected for collecting microorganisms and the like includes, for example, the back of a refrigerator handle. For this reason, it is preferable that the equipment used for the collection can be simply used by using a material having excellent operability such as a length.

  In order to solve the above-described problems, the present invention efficiently collects microorganisms and genes that can be easily collected from a surface to be inspected, microorganisms and genes, and microorganisms or genes collected by the collection method. It is an object of the present invention to provide a microorganism and gene extraction method that can be easily extracted.

<1> A liquid medium is used for the absorber of the collector material, using a collector material provided with an absorber, and a rod-shaped member that can separate the tip portion and the carrier portion provided with the absorber at one end. A first step of absorbing;
A second step of collecting microorganisms or genes on the surface to be inspected by bringing the absorber of the collection device into contact with the surface to be inspected;
Using a first container that has an opening at one end and can be sealed by attaching a lid to the opening, the collector is inserted into the opening of the first container from the absorber side, and inserted After separating the tip portion of the rod-shaped member and the carrier portion, the carrier portion is taken out from the first container, and the first container in which the tip portion is accommodated so that the insertion direction is maintained is removed. A third step of sealing;
Using a second container that has an opening at one end and can be sealed by attaching a lid to the opening, the tip of the collector is connected to the second side from the side opposite to the absorber. The fourth container is inserted from the first container into the second container so as to be inserted into the opening of the container, and the second container in which the tip is stored is sealed so that the insertion direction is maintained. And the process of
A microorganism and a method for collecting genes.
<2> The microorganism and gene collection method according to <1>, wherein the liquid medium has a volume of 20 μL to 100 μL.
<3> The microorganism and gene collection method according to <1> or <2>, wherein the liquid medium is stored in the first container in the first step.
<4> The microorganism and gene collection method according to any one of <1> to <3>, wherein the length of the exposed portion of the tip of the collection device is 5 mm to 10 mm.
<5> The microorganism and gene collection method according to any one of <1> to <4>, wherein the tip portion and the support portion can be separated by breaking the collection device.
<6> The microorganism and gene collection method according to any one of <1> to <5>, wherein the collection device is a cotton swab capable of separating a tip portion and a support portion.
<7> A microorganism and gene extraction method using the microorganism and gene collection method according to any one of <1> to <6>,
The second container obtained in the fourth step is attached to a centrifuge so that the absorber side of the tip portion housed in the second container is on the centrifugal shaft side, and then centrifuged. A microorganism and gene extraction method comprising a fifth step of performing the steps.

  According to the present invention, a microorganism that can easily collect a microorganism or gene from the surface to be inspected, a method for collecting the gene, a microorganism that can efficiently and simply extract the microorganism or gene collected by the collection method, and A method for extracting a gene can be provided.

It is sectional drawing for demonstrating the relationship between the front-end | tip part side edge part of the absorber in a cotton swab, and the support part side edge part of a front-end | tip part. It is the schematic which shows the flow of the 1st and 2nd process of this embodiment. It is the schematic which shows the flow of the 3rd process of this embodiment. It is the schematic which shows the flow of the 4th process of this embodiment. (5-a) is a side view schematically showing a centrifuge used in the fifth step of the present embodiment, and (5-b) shows the centrifuge used in the fifth step and the second step. It is a top view which shows the relationship with a container.

  Hereinafter, the contents of the present invention will be described in detail using embodiments. However, the following embodiments are examples, and the present invention is not limited to these embodiments.

The microorganism and gene collection method of the present embodiment (hereinafter sometimes simply referred to as “collection method of the present embodiment”) is a method including the following first to fourth steps.
(First step)
A step of causing the absorber of the collector material to absorb a liquid medium, using a collector member including an absorber and a rod-shaped member capable of separating the tip portion and the support portion provided at one end of the absorber. (Second step)
A step of collecting microorganisms or genes on the surface to be inspected by bringing the absorber of the collecting device into contact with the surface to be inspected (third step)
Using a first container that has an opening at one end and can be sealed by attaching a lid to the opening, the collector is inserted into the opening of the first container from the absorber side, and inserted After separating the tip portion of the rod-shaped member and the carrier portion, the carrier portion is taken out from the first container, and the first container in which the tip portion is accommodated so that the insertion direction is maintained is removed. Sealing step (fourth step)
Using a second container that has an opening at one end and can be sealed by attaching a lid to the opening, the tip of the collector is connected to the second side from the side opposite to the absorber. Inserting the first container from the first container into the second container so as to be inserted into the opening of the container, and sealing the second container containing the tip so that the insertion direction is maintained

The microorganism and gene extraction method of the present embodiment (hereinafter sometimes simply referred to as “extraction method of the present embodiment”) is the second container obtained through the first to fourth steps described above. Is used to extract microorganisms and the like by the following fifth step.
(Fifth step)
The second container obtained in the fourth step is attached to a centrifuge so that the absorber side of the tip portion housed in the second container is on the centrifugal shaft side, and then centrifuged. Process

According to the collection method of this embodiment, microorganisms and the like present on the surface to be inspected can be easily collected and sealed. In particular, according to the collection method of the present embodiment, the collection device that has collected microorganisms and the like is separated to remove the carrier, and only the tip is stored in the first container and sealed, which is convenient. Microbes can be collected with high sealing properties. For this reason, even in stores, factories, etc., it is possible to easily collect microorganisms and the like on the surface to be inspected without requiring special knowledge by employees.
Moreover, according to the extraction method of this embodiment, it is possible to extract efficiently the microorganisms or gene collected by the absorber of the collection device material with a small amount of liquid medium by centrifugation. For this reason, the obtained collected liquid can be subjected to microscopic examination such as direct observation with a microscope or the like, genetic examination, immunological examination or culture examination without going through a concentration step.

Here, the microorganisms (including bacteria) or genes to be examined in this embodiment include, for example, protozoa such as Cristosporidium and dysentery amoeba, fungi such as Candida and yeast, mold, enterohemorrhagic Escherichia coli, intestinal invasion Escherichia coli, Shigella, Toxigenic Escherichia coli, Salmonella, Listeria, Vibrio, Campylobacter, Staphylococcus aureus, Clostridium perfringens, Clostridium botulinum, Bacillus cereus, etc. It includes genes such as bacteria, norovirus, sapovirus, rotavirus, adenovirus, astrovirus and other viruses such as diarrhea virus, enterovirus, hepatitis virus, coxsackie virus, and other traces of microorganisms.
The collection method and extraction method of the present embodiment (hereinafter, these may be collectively referred to as “method of the present embodiment”) are capable of collecting and extracting microorganisms and the like with a particularly small amount of liquid medium. Therefore, it is preferably used as a method for collecting and extracting pathogenic viruses and pathogenic bacteria (for example, norovirus, enterohemorrhagic Escherichia coli, Campylobacter, etc.) that can be infected with a small number of bacteria, microorganisms, or genes, or the number of viruses. be able to. For example, norovirus may be infected even in an abundance such as 10-100. For this reason, if the collected liquid is used for inspection without being concentrated, it may be difficult to say that it is completely infectious even if the test result is “negative”. On the other hand, when an undiluted collection liquid is used, if the test result is “negative”, there is a high possibility that there is no infection and the reliability of the test result is high.

[The collection method of this embodiment]
As described above, the collection method of the present embodiment is a method including first to fourth steps. Hereinafter, the first to fourth steps will be described.

(First step)
In the method according to the present embodiment, the first step uses a collecting device including an absorber, and a rod-shaped member that can separate the tip and the support provided with the absorber at one end. In this step, the liquid medium is absorbed by the absorber of the collector. By absorbing the liquid medium in the absorber in the step, microorganisms and the like existing on the surface to be inspected can be collected in the absorber when the surface to be inspected is wiped by the absorber.

-Collector equipment-
In this embodiment, the collector device includes an absorber and a rod-like member, and the absorber is provided at one end of the rod-like member. The collector material in the present embodiment can be separated into the tip portion and the support portion provided with the absorber by separating the rod-like member into two. Moreover, since the collector material in this embodiment uses the rod-shaped member, it is excellent in the operativity in the case of wiping off.

  The absorber is a member that can collect and maintain microorganisms and the like wiped off by absorbing the liquid medium. Moreover, the microorganisms etc. which were maintained in the absorber can be extracted with a liquid medium by performing centrifugation etc. to an absorber. The material constituting the absorber is not particularly limited as long as it can absorb a liquid medium described later and collect microorganisms and the like on the surface to be inspected. Examples of materials that can be used for the absorber include cotton balls, sponges, and gauze. Further, the size and shape of the absorber are not particularly limited, and are appropriately selected according to the size of first and second containers (for example, a microtube for a centrifuge used for microbiological examination) described later. can do. However, according to the method of the present embodiment, from the viewpoint of collecting microorganisms and the like with a small amount of liquid medium, for example, the size of the absorber is determined so that the amount of liquid absorbed is 20 to 100 μL. can do.

The rod-shaped member is a member that functions as a support member for the absorber. Further, the rod-shaped member is designed so as to be separated into at least a tip end portion and a carrying portion. The means for separating the tip portion and the carrier portion is not particularly limited.For example, when the strength of the separation portion between the tip portion and the carrier portion is made lower than other portions and a force is applied from the outside. It can design so that the said isolation | separation site | part can be separated, such as breaking. In order to reduce the strength of the separation part, for example, there are methods such as notching the rod-like member or making the diameter of the part corresponding to the separation part thinner than other parts.
The material used for the rod-shaped member is not particularly limited as long as it has a certain degree of rigidity and does not absorb liquid, and examples thereof include a plastic material such as polystyrene. Further, the tip portion and the carrying portion may be formed of the same material or different materials. In addition, the size (full length) of the tip of the rod-shaped member is not particularly limited. For example, the size can be attached to the opening after being accommodated in each container in the third and fourth steps described later. (That is, shorter than the total length of each container), and when the collected liquid is extracted in the fifth step, the position of the absorber in the second container is lower than the liquid level of the collected liquid. It is preferable that the length be located on the opening side of the container. Further, the size of the support portion of the rod-shaped member is not particularly limited, but the length is about 4 cm to 18 cm in order to improve the visibility after separation and the operability at the time of collecting microorganisms (wiping off). Preferably 6 to 13 cm.

  As the collector device, for example, a cotton swab in which an absorbent body is formed of a cotton ball and a rod-shaped member is formed of plastic or the like, which can separate a tip portion and a supporting portion, can be used. When the collecting device is a cotton swab, the size of the absorbent body (cotton ball) is, for example, 3 mm to 10 mm in diameter and 8 mm to 15 mm in length, and the size of the entire rod-shaped member is, for example, 2 mm to 5 mm in diameter and 6 cm in length. It can be set to ˜20 cm. At this time, the length of the exposed portion of the tip portion (distance from the tip portion side end portion of the absorber to the end portion of the tip portion on the carrier portion side) can be set to, for example, 5 mm to 10 mm. The length of the exposed portion of the tip affects the effect of suppressing the contact between the collection liquid extracted on the tip side of the microtube and the cotton ball at the time of centrifugation in the fifth step to be described later. To get. For this reason, the above-mentioned “end part on the tip side of the absorber” is based on the part located on the most tip side of the absorber.

  For example, a common cotton swab is configured such that the tip of a rod-shaped member is inserted into a cotton ball, and a part of the cotton ball covers the surface of the rod-shaped member. The relationship between the tip part side end part of the cotton ball (absorber) of the cotton swab and the carrying part side end part of the tip part will be described with reference to FIG. FIG. 1 is a cross-sectional view for explaining the relationship between an end portion side end portion of an absorbent body and an end portion on a support portion side of a cotton swab. As shown in FIG. 1, when the tip part 24 of the rod-shaped member is inserted into the cotton ball 22, a part of the tip part 24 is covered with a part of the cotton ball 22. In this case, a portion of the end of the fiber of the cotton ball 22 (so-called “brow”) that is located closest to the tip is the end of the cotton ball 22 (absorbent body) (the dotted line E2 in FIG. 1). ). That is, in FIG. 1, the distance from the tip of the cotton ball 22 (dotted line E1 in FIG. 1) to the dotted line E2 is “the length of the cotton ball”. Similarly, in FIG. 1, the distance from the tip end side end portion (dotted line E2) of the cotton ball 22 to the carrying portion side end portion (dotted line E3 in FIG. 1) of the tip end portion 24 is "the exposed portion of the tip end portion". Length ”.

-Liquid medium-
The liquid medium is a material that assists the absorber in collecting microorganisms and the like, and an aqueous solution generally called a buffer solution or a diluent can be used. As the liquid medium, for example, a buffer solution such as physiological saline or phosphate buffered saline (PBS), water, or the like can be used. Further, the liquid medium is not necessarily in a liquid state, and may be, for example, in a gel form as long as it is a medium that can assist microorganisms and the like to be collected by the absorber.

  In the first step, the liquid medium is absorbed by the absorber of the collector. The volume of the liquid medium absorbed by the absorber is preferably 20 μL to 100 μL, and preferably 30 μL to 40 μL from the viewpoint of extracting the collected liquid by centrifugal force and efficiently detecting microorganisms without going through a concentration step. Is more preferable. In the first step, the liquid medium can be used in a first container, which will be described later, stored in the container in advance. In this case, the lid can be removed from the first container, and then the absorber of the collector can be inserted into the container to absorb the liquid medium. In addition, in the liquid medium, from the viewpoint of promoting the migration of the microorganisms collected in the second step or ensuring the survival of the collected microorganisms, surfactants, nutrients of the microorganisms, gelling agents, salts, and the like are added. It may be added.

(Second step)
The second step is a step of collecting microorganisms or genes on the surface to be inspected by bringing the absorber of the collection device into contact with the surface to be inspected. In the process, microorganisms and the like are collected in the absorber. The method for bringing the absorber into contact with the surface to be inspected is not particularly limited. For example, a method for wiping the surface to be inspected with the absorber is given as an example.

(Third step)
The third step uses a first container that has an opening at one end and can be sealed by attaching a lid to the opening, and the collector is opened from the absorber side to the opening of the first container. Inserted into the first part, and after the insertion, the tip part of the rod-shaped member and the carrier part are separated, the carrier part is taken out from the first container, and the tip part is stored so that the insertion direction is maintained. A step of sealing the first container; This step is a step of sealing only the tip of the collection device provided with the absorber in the first container after the microorganisms on the surface to be inspected are collected in the absorber in the second step.

The collection device collects microorganisms on the surface to be inspected in the absorber in the second step, and then is inserted into the opening of the first container from the absorber side, and then separated into a tip portion and a support portion. The The method for separating the tip portion and the support portion is not particularly limited. For example, the tip portion and the support portion are separated by using the wall surface of the first container and folding the collector material using the lever principle. Can be separated. The carrier part separated from the tip part is taken out from the first container, and then the lid is attached to the opening part to seal the first container. As described above, according to the method of the present embodiment, without directly touching the absorber and its peripheral portion, the carrier portion can be removed and only the tip portion provided with the absorber can be sealed in the first container. it can. For this reason, it is possible to effectively prevent the microorganisms and the like collected in the first container from being mixed with microorganisms or the like derived from workers (mainly fingers) performing the first, second, and third steps. be able to.
Moreover, the collector member (tip portion) stored in the first container is the other end side (that is, the closed side) of the first container on the side where the absorber is provided, and even after storage. The direction (insertion direction) in which the collector device is inserted is maintained in the first container.

-First container-
The first container has an opening at one end, and is a container that can be sealed by attaching a lid to the opening. Further, the other end of the first container is closed with a wall surface or the like. For example, when the other end is set downward, the liquid in the container is accumulated. The size and shape of the first container are not particularly limited, and for example, a known sealable container such as a microtube used in a centrifuge can be appropriately used.
The opening of the first container is configured so that a lid can be attached, and the first container is sealed by attaching the lid to the opening. The lid can be attached to the opening by applying known means such as screwing or pressure bonding means.
The material of the first container is not particularly limited, and a known material such as a material used for the microtube can be appropriately used. For example, a colored material can be used. Moreover, after the front-end | tip part of a rod-shaped member is accommodated, the 1st container needs to be the shape by which the insertion direction is maintained. Examples of such a shape include a cylindrical body in which the inner diameter of the first container is smaller than the total length (including the absorber) of the tip. The size of the first container can be, for example, 8 mm to 15 mm in diameter and 30 mm to 50 mm in length, and a microtube for 2.5 mL, 2 mL, or 1.5 mL can be used in terms of volume. Further, as described above, the first container may be one in which the above-described liquid medium is stored in advance, for example, 20 μL to 100 μL, preferably 30 μL to 40 μL.

(Fourth process)
The fourth step uses a second container that has an opening at one end and can be sealed by attaching a lid to the opening, and the tip of the collector is opposite to the absorber. The second container is inserted from the first container into the second container so as to be inserted into the opening of the second container from the side, and the tip is stored so that the insertion direction is maintained. It is a process of sealing the container. This step is a step in which the collector device sealed in the first container in the third step is transferred to the second container with the insertion direction reversed and then sealed.

In this step, the collector device taken out from the first container is inserted into the opening of the second container from the side opposite to the side where the absorber is provided. Since the absorber side of the collector material is located on the side opposite to the opening (that is, the other end side) in the first container, the opening of the second container remains in the state of being taken out from the opening of the first container. If the collector is inserted into the part, the collector (tip) is inserted into the opening of the second container from the side opposite to the side where the absorber is provided.
After being inserted into the second container, the collector material is sealed so that the direction in which the collector material is inserted (insertion direction) is maintained in the second container even after storage.

-Second container-
The second container has an opening at one end and can be sealed by attaching a lid to the opening, and the same container as the first container can be used.

[Extraction method of this embodiment]
As described above, the extraction method of the present embodiment is a method of extracting microorganisms and the like by the fifth step using the second container obtained through the first to fourth steps.

(Fifth step)
In the fifth step, the second container obtained in the fourth step is separated from the centrifuge so that the absorber side of the tip portion accommodated in the second container is on the centrifugal shaft side. It is the process of attaching to and performing centrifugation. In this step, the second container is attached to the centrifuge so that the absorber side of the stored tip is on the centrifuge shaft side, and the microorganisms collected in the absorber are collected by centrifugation. The liquid medium (collected liquid) containing is extracted to the other end side (opposite side of the opening) of the second container. The collector in the 2nd container which passed through the 4th process has an absorber located in the opening part side, and the other end side serves as a tip part. Thus, since the gap can be provided between the wall surface on the other end side (the side opposite to the opening) in the second container and the absorber due to the presence of the tip, the other end of the second container Suppressing the contact between the absorbent and the collected liquid when the collected liquid is accumulated on the other end side of the second container by centrifuging by setting the side to be outside the rotation axis be able to.

-Centrifuge-
The centrifuge that can be used in this step can install the second container around the rotation axis, the rotation rotor rotates around the rotation axis by the drive mechanism, and the microorganisms and the like from the absorbent in the second container Can be centrifuged. The centrifuge is not particularly limited, and for example, a commercially available product such as a desktop small microcentrifuge can be appropriately selected and used. Centrifugation conditions are not particularly limited, and can be appropriately set according to the amount of sample used and the apparatus. Centrifugation can be performed by low-speed centrifugation (for example, 3 to 10 seconds at a centrifugal force of 1000 to 3000 [× g]).

-Collecting liquid-
Since the collected liquid extracted in this step is a highly efficient extraction of microorganisms and liquid medium collected in the absorber, direct observation with a microscope or the like without performing a concentration step, It can be used for microbial tests such as genetic tests, immunological tests or culture tests.

[First Embodiment]
Hereinafter, the method of this embodiment will be specifically described with reference to the drawings.
FIG. 2 is a schematic diagram showing the flow of the first and second steps of the present embodiment. As shown in FIG. 2, the microtube 10 (first container) includes a tube body 2 and a lid body 4. An opening 8 is provided at one end (upper side of the drawing) of the tube body 2, and the microtube 10 can be sealed by attaching the lid 4 to the opening 8. As shown in FIG. 2, the tube body 2 has an opening 8 at one end, and the other end (tip: the lower side on the paper surface) is closed by a wall surface. In addition, the tube body 2 has a portion having a constant length and the same diameter from the opening 8 side toward the distal end side, and further, the diameter is reduced toward the distal end side to become a tapered shape. Has a site. The other end of the tube main body 2 forms the tip of the microtube 10 and can serve as a liquid reservoir inside. In FIG. 2 (1-a), the buffer solution 6 (liquid medium) is stored inside the microtube 10. Note that the buffer solution 6 in the microtube 10 does not necessarily have to be accumulated at the tip side in a normal state, and the buffer solution 6 is collected at the tip by shaking the microtube 10 with the opening 8 side as a fulcrum at the time of use. Can do.

  As shown in (1-b) of FIG. 2, first, a cotton swab 20 (collector) having a cotton ball 22 (absorber) and a rod-shaped member that can separate the tip 24 and the support 26 is used. It inserts in the microtube 10 and makes the cotton ball 22 absorb the buffer solution 6 (first step). The cotton swab 20 is provided with a notch 28 as a means for separating the tip 24 and the carrier 26.

  Next, as shown in FIG. 2 (2), the cotton swab 20 having absorbed the buffer solution 6 is pulled out of the microtube 10, and then the surface 32 to be inspected is wiped so that the cotton ball 22 of the cotton swab 20 is in contact with it. Microorganisms or genes on the surface 32 to be inspected are collected in the cotton ball 22 (second step).

  A 3rd process is demonstrated using FIG. FIG. 3 is a schematic diagram showing the flow of the third step of the present embodiment. As shown to (3-a) of FIG. 3, the cotton swab 20 which collected microorganisms etc. is inserted in the opening part 8 of the microtube 10 from the cotton ball 22 side. Next, as shown in FIG. 3 (3-b), the cotton swab 20 is folded using the wall surface of the microtube 10 so that a force is applied to the notch 28, and the tip 24 and the carrier 26 are separated. To do. Thereafter, as shown in FIG. 3 (3-c), only the carrier 26 is taken out from the microtube 10, the tip 24 is left, the lid 4 is attached, and the microtube 10 is sealed. At this time, in the microtube 10, the insertion direction of the distal end portion 24 is maintained so that the cotton ball 22 is positioned on the other end side (the lower side of the drawing) of the microtube 10. For example, if the place where the microorganisms are collected is different from the place where the microorganisms are examined, the microtube 10 sealed in the third step is used as an inspection center where the microorganisms are collected from the place where the microorganisms are collected. Can be transported. In this embodiment, since the microtube 10 is excellent in portability and sealing performance, the microtube 10 can be mailed to an inspection center or the like at a low cost, for example, in an envelope.

  A 4th process is demonstrated using FIG. FIG. 4 is a schematic diagram showing the flow of the fourth step of the present embodiment. In a 4th process, the front-end | tip part 24 sealed by the microtube 10 is moved to the microtube 30, and the microtube 30 is sealed after that. In the present embodiment, the microtube 10 and the microtube 30 having the same shape and size are used. However, the microtube 10 and the microtube 30 may have different shapes or sizes.

  As shown to (4-a) of FIG. 4, in the 4th process of this embodiment, the microtube 30 (2nd container) is used, the cover body 34 is remove | desorbed from the microtube 30, and also a microtube. The lid 4 is detached from 10, and the tip 24 of the cotton swab accommodated in the microtube 10 is inserted into the microtube 30 through the opening 36 from the microtube 10. At this time, the tip 24 is inserted into the microtube 30 by sliding down the wall surface of the microtube 10 as indicated by an arrow in the figure. For this reason, the transition from the microtube 10 to the microtube 30 can be performed without the tip 24 of the swab touching a human finger or the like. Further, in the tip part 24 of the swab, the cotton ball 22 becomes the opening 36 side (upper side of the paper surface) of the microtube 30 and the insertion direction is maintained. Thereafter, the lid 34 is attached, and the microtube 30 is sealed while the tip part 24 of the cotton swab is stored.

  Next, the fifth step will be described. In FIG. 5, (5-a) is a side view schematically showing a centrifuge used in the fifth step of the present embodiment, and (5-b) is a centrifuge used in the fifth step. It is a top view which shows the relationship between a 2nd container. Although not described in detail, as shown in FIG. 5 (5-a), the centrifuge 40 includes a rotating rotor 42 that rotates about a rotating shaft (centrifugal shaft) R indicated by a one-dot chain line. A plurality of microtubes 30 are installed on the rotating rotor 42.

  As shown in FIG. 5 (5-b), the plurality of microtubes 30 are the microtubes 30 obtained by the fourth step, and the cotton balls 22 of the tip part 24 of the cotton swabs housed in the container. The centrifuge 40 is mounted so that the side is the rotation axis R side. In the present embodiment, eight microtubes 30 are installed in the centrifuge 40, but the number of microtubes 30 is not particularly limited.

  In the fifth step, the rotating rotor 42 rotates at high speed around the rotation axis R by a drive mechanism (not shown), and the collected liquid containing microorganisms and the like is extracted from the cotton balls 22 into the microtube 30 by centrifugal force. The After centrifugation, the collected liquid extracted from the cotton ball 22 is collected on the tip side of the microtube 30. At this time, due to the presence of the distal end portion 24, it is possible to suppress contact between the collected liquid extracted on the distal end side of the microtube 30 and the cotton ball 22.

  According to the method of the present embodiment, for the extraction of microorganisms and the like collected in the second step, the insertion direction of the tip portion 24 in the third step and the tip portion 24 in the fourth step (the cotton balls 22 are By reversing the (positioning direction), the collected liquid can be efficiently extracted from the cotton ball 22 (absorber) in the fifth step. For this reason, the collection liquid extracted in this step can be used for microscopic examinations such as direct observation with a microscope or the like, genetic examination, immunological examination or culture examination without performing a concentration process.

  As mentioned above, although the collection method of this invention and the extraction method were demonstrated using the Example, this invention is not limited to this.

2 tube body, 4 lid, 6 buffer solution (liquid medium), 8 opening, 10 microtube (first container), 20 cotton swab (collector), 22 cotton ball (absorber), 24 tip, 26 carrying part, 28 notch part, 30 microtube (second container), 32 surface to be inspected, 34 lid, 36 opening, 40 centrifuge, 42 rotating rotor

Claims (7)

First, a collector member including an absorber and a rod-shaped member capable of separating a tip portion and a carrier portion provided at one end of the absorber is used to cause the absorber of the collector member to absorb a liquid medium. 1 process,
A second step of collecting microorganisms or genes on the surface to be inspected by bringing the absorber of the collection device into contact with the surface to be inspected;
Using a first container that has an opening at one end and can be sealed by attaching a lid to the opening, the collector is inserted into the opening of the first container from the absorber side, and inserted After separating the tip portion of the rod-shaped member and the carrier portion, the carrier portion is taken out from the first container, and the first container in which the tip portion is accommodated so that the insertion direction is maintained is removed. A third step of sealing;
Using a second container that has an opening at one end and can be sealed by attaching a lid to the opening, the tip of the collector is connected to the second side from the side opposite to the absorber. The fourth container is inserted from the first container into the second container so as to be inserted into the opening of the container, and the second container in which the tip is stored is sealed so that the insertion direction is maintained. And the process of
A microorganism and a method for collecting genes.   The method for collecting microorganisms and genes according to claim 1, wherein the volume of the liquid medium is 20 μL to 100 μL.   The method for collecting microorganisms and genes according to claim 1 or 2, wherein in the first step, the liquid medium is stored in the first container.   The method for collecting microorganisms and genes according to any one of claims 1 to 3, wherein the length of the exposed portion of the tip of the collection device is 5 mm to 10 mm.   The method for collecting microorganisms and genes according to any one of claims 1 to 4, wherein the tip and the carrier can be separated by breaking the collecting device.   The method for collecting microorganisms and genes according to any one of claims 1 to 5, wherein the collecting device is a cotton swab capable of separating a tip portion and a supporting portion. A microorganism and gene extraction method using the microorganism and gene collection method according to any one of claims 1 to 6,
The second container obtained in the fourth step is attached to a centrifuge so that the absorber side of the tip portion housed in the second container is on the centrifugal shaft side, and then centrifuged. A microorganism and gene extraction method comprising a fifth step of performing the steps.