JP2008200028A - Tube container for reaction or detection, and reagent kit comprising the container - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a tube container preventing fly-up of drying agents, caused when throwing a specimen sample therein, decreasing processing procedures of an experimenter, quickly and well performing nucleic acid amplification and preventing decrease of analytical accuracy. <P>SOLUTION: The tube container is provided by integrally forming a reaction tube, a lid sealing an opening part of the upper surface of the reaction tube, and a connecting part connecting the upper part of the reaction tube with the lid, wherein the reaction tube has a cylindrical barrel ranging from the opening part, a tapered part formed to be tapered from the barrel, and a bottom part formed to close the front end of the tapered part, and integrally provided with the reaction tube a reagent supporting part capable of holding a carrier supporting inside the tapered part a reagent or a carrier supporting the reagent, at the position thereof by drying or freeze-drying. And the reagent kit includes the tube container and the drying reagent or the carrier supporting the reagent. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a tube container suitable for microanalysis of a sample and a reagent kit including the container.

Nucleic acid amplification methods are used to efficiently analyze particularly target nucleic acids in genetic analysis of biological samples such as blood and urine, foods, and environmental water.
When carrying out the nucleic acid amplification method, since the nucleic acid obtained as a sample is very small, the reaction itself is carried out in a very small amount of solution, usually in a solution of several tens of μL.
For this reason, the nucleic acid amplification reaction is usually carried out in a reaction tube with a small capacity lid, and various reaction tubes have been proposed.

For example, a plastic reaction container containing a small amount of liquid in which a lid and a reaction container are integrated has been proposed. The lid can be easily opened and closed, and there is almost no risk of being opened by the internal pressure of the container. In order to securely hold the lid, the fitting property between the lid and the container opening, and the lid and the coupling body have been studied (see Patent Document 1).
In addition, as a reaction tube that reduces the frequency of opening and closing the lid and increases the number of samples that can be processed at one time, a reaction tube having a container-integrated partition wall in the reaction container has been proposed (see Patent Document 2). .
Furthermore, in an integrated reaction vessel having a lid and a container opening, there is disclosed a container in which the lid has an extremely thin film, and two folding grooves are provided in a folding hinge of the lid and the container connecting portion. (See Patent Document 3).
JP-A-60-183362 Japanese Utility Model Publication No. 7-34935 JP-T 9-504690

The tube containers for nucleic acid amplification that have been studied so far have been designed to improve the openability of the lid, the adhesion between the lid and the container, or the efficiency of sample processing.
When nucleic acid amplification is carried out using the above-mentioned container, for example, a nucleic acid amplification reagent is carried on a carrier such as a non-woven fabric and dried, and a nucleic acid amplification reaction is performed by introducing a detection sample toward the reagent. The means to do is adopted. However, in this case, when the sample is charged, the dry reagent does not exist in the predetermined reaction zone or detection zone of the tube container, and soars upward and stays in that position (this is called “swelling”). The dry reagent may adhere to various parts other than the original reaction zone and detection zone of the inner wall of the container. At this time, it is not only necessary for the experimenter to temporarily stop the analysis procedure such as stopping the operation of the detection analyzer and to drop the dry reagent to the lower part of the container, but also nucleic acid amplification cannot be performed quickly and satisfactorily. There is a problem that the detection accuracy is insufficient and the analysis accuracy is remarkably lowered (sometimes analysis is impossible).
Further, this “swelling” phenomenon has been a problem when it is stored, transported, or used in a reagent kit in which a carrier having a nucleic acid amplification reagent previously loaded in the lower part of the tube container is inserted.

As a result of diligent research to solve the above problems, the present inventors have provided a reagent holding part at the bottom of a tube container that can store a small amount of liquid sample, and thus, after the sample for specimen has been introduced so far The present inventors have found that the rise of the dry reagent that has occurred can be reduced.
That is, the present invention is a tube container for performing reaction or detection of a target substance in a small amount of a sample, which is a reaction tube, a lid portion that seals an opening on the upper surface, an upper portion of the reaction tube, and the The reaction tube is integrally formed with a joint portion that joins the lid portion, and the reaction tube includes a cylindrical barrel portion that is continuous from the opening portion, a tapered portion that is tapered from the trunk portion, Reacting reagent holding part which has a bottom part formed so as to close the tip, and can hold the reagent or the carrier carrying the reagent in its position by drying or freeze-drying the reagent The present invention relates to a tube container provided integrally with a tube.

In a preferred aspect of the present invention, the reagent holding part is composed of a projecting part projecting from the inner wall surface of the tapered part toward the center of the reaction tube, and the projecting part preferably extends over the inner periphery of the tapered part. Thus, it is desirable that 2 to 10 pieces, more preferably 3 to 6 pieces are provided to face each other. Moreover, it is desirable that the overhanging portion extends from the lower portion of the tapered portion to the bottom portion.
In the present invention, the overhang portion may be a rib.

  Furthermore, in another preferred aspect of the present invention, the reagent holding portion is formed of a recess formed on the inner wall of the tapered portion over the inner periphery of the tapered portion, and the recessed portions are preferably opposed to each other. It is desirable to provide 10 to 10, more preferably 3 to 6. Furthermore, it is desirable that the recess extends from the lower part of the taper to the bottom.

  In the tube container of the present invention, it is desirable that a display area having a larger area for displaying the contents of the container is provided on the lid.

  The present invention further relates to a reagent kit comprising the aforementioned tube container and a dry reagent held in the reagent holding part of the tube container or a carrier capable of carrying the dry reagent.

The tube container according to the present invention is provided with a reagent holding section that can hold a reagent or a carrier carrying the reagent in its position by drying or freeze-drying the reagent at the lower part of the container (near a predetermined reaction zone or detection zone). ing.
As an example of a method for holding a reagent or a carrier carrying a reagent in the reagent holding portion of the tube container, the following method can be exemplified.
(A) A support made of non-woven fabric or the like is adhered to the reagent holder of the tube container, and then a liquid reagent is poured into the support so as to be immersed (to serve as a carrier carrying the reagent) and dried under reduced pressure. Let
(B) A liquid reagent is put into a tube container and dried or freeze-dried in the container.
By the above-mentioned method, the dried reagent adheres to the overhanging portion or the concave portion provided in the reagent holding portion, or is sandwiched between the overhanging portion / recessed concave / convex shape, or stays so as to be bridged. It becomes impossible to leave the holding part.
For this reason, for example, even when the tube container is tilted or stirred with a stirrer, the dried reagent or the like is then moved from the reagent holding unit, and the predetermined reaction zone of the tube container is moved. Or it can prevent adhering to inner walls other than a detection area.

  In addition, when the reaction or detection of a target substance in a small amount of sample is performed using the tube container of the present invention, the reagent or sample does not rise in the container and remains in a predetermined reaction area or detection area at the bottom of the container. Thus, it is possible to reduce the labor (in the process of dropping the dry reagent to the lower part of the container) in the analysis procedure caused by the “float”, and to prevent problems such as poor analysis accuracy and inability to analyze.

  Moreover, since the reagent kit of the present invention uses the tube container of the present invention and can hold the reagent in a dry state, the reagent kit can be quickly and accurately implemented when the reaction or detection of the target substance in the sample is necessary.

  A tube container for reaction or detection of a target substance in a small amount of a sample according to the present invention is a reaction tube, a lid portion that seals an opening on the upper surface, and an upper portion of the reaction tube and the lid portion. The joint part is integrally molded.

The tube container of the present invention is preferably made of plastic.
Plastic can be molded into a tube container very easily and at once by a known and commonly used method such as injection molding, and can be mass-produced at low cost.
The tube container used in the present invention is manufactured from a resin appropriately selected from polyethylene resins, polypropylene resins, other polyolefin resins, polyester resins, fluorine resins, silicon resins, and the like.
In addition, the tube container of the present invention can be adjusted by appropriately adjusting the diameter of the reaction tube, the size and thickness of the container according to the reaction / detection mechanism of the target substance and the size of the apparatus used for the reaction / detection. Good.

[Cover]
In the tube container of the present invention, the lid portion has a function of sealing the opening on the upper surface of the reaction tube, and is integrally formed with the reaction tube and a joint portion described later.
The lid portion is adapted to the opening of the reaction tube and has a cylindrical seal portion formed so as to obtain a suitable hermetic seal against the inner wall surface of the tube that extends below the periphery of the opening. It is formed over the entire outer periphery at the end of the cylindrical seal portion, and has an outer peripheral portion formed so as to be in close contact with the edge of the reaction tube and to obtain a suitable hermetic seal against it. .
The cylindrical seal portion is formed so that the adhesion is enhanced when fitted to the opening of the reaction tube, and is easily removed when removed from the opening, if necessary. The outer periphery of the end portion farthest from the lid main body is formed slightly larger than the outer periphery of the seal portion closest to the lid main body.

In addition, the lid portion is provided with a display area with a larger area for facilitating opening and closing of the lid portion (removal of the seal portion of the lid portion from the reaction tube opening) and showing the contents of the container. Is preferred. The display portion is formed continuously from the outer peripheral portion of the lid portion.
When viewed from the upper surface of the tube container, the shape of the display portion may be various, such as a substantially square shape or a substantially triangular shape, but preferably has a substantially square shape for the above purpose.

[Joint part]
The tube container of this invention WHEREIN: The said junction part is integrally molded with the said reaction tube and the said cover part. The joint portion is provided so as to connect the upper portion of the reaction tube and the lid portion to prevent the lid portion from being lost, and the lid portion is easy to handle when sealing the opening of the reaction tube. It has the function of improving.

The bonding portion of the joint with the reaction tube side may be the peripheral edge of the opening of the reaction tube or the outer wall surface above the trunk of the reaction tube.
The joining portion is formed so that the sealing portion of the lid portion can be easily inserted from the opening portion of the reaction tube, and is easily bent with flexibility at approximately the center so that it does not easily come off after insertion. It is desirable that If it is not formed so as to be easily bent at the approximate center, a force is exerted to return the joint from the bent state over time, and the adhesion between the lid and the reaction tube opening is reduced. In addition, in some cases, the lid is removed from the reaction tube opening. For this reason, the joint portion is formed with a structure having a hinge at the center or a curved structure, or formed so as to change the thickness or width so that it can be easily bent at the center of the joint portion. The
Moreover, although this junction part is normally formed in one strip shape, you may form in two or more string shape.

[Reaction tube]
In the tube container according to the present invention, the reaction tube includes a cylindrical barrel portion that is continuous with the opening, a tapered portion that is tapered from the barrel portion, and a bottom portion that is formed so as to close a tip of the tapered portion. It has.
As described above, the opening portion has an edge portion that can be easily sealed by the lid portion. When the reaction tube is viewed from the upper surface, generally, the edge portion is formed so as to protrude slightly outward from the outer periphery of the cylindrical body portion, that is, has a larger area than the cross section of the cylindrical body portion.

  In the tube container, it is preferable that the inner surface has a smooth surface, but the outer surface is formed for the purpose of improving the strength of the container or corresponding to the internal shape of the detection device to be used. A groove or the like may be formed in the longitudinal direction or the circumferential direction.

  In the taper formed so as to be tapered from the body part, the angle of the taper of the inner surface is appropriately selected according to the internal shape of the detection device to be used. Preferably, however, if the dried sample rises It is desirable for the taper angle to be large because it is easy to fall to a predetermined position at the bottom of the container, and when the liquid sample is introduced, even if the sample adheres to the wall surface, it tends to fall to the predetermined position.

  In the tube container, examples of the shape of the inner surface of the body part include a cylindrical shape such as a cylinder (regular cylinder, elliptical column, etc.) and a prism (triangular column, square column, etc.), and the inner surface of the tapered portion. Examples of the shape include a cone shape (conical shape, pyramid shape (triangular pyramid, quadrangular pyramid, etc.)) and the like. The inner surface of the barrel and the inner surface of the tapered portion may be any combination of shapes, but when the dried sample soars, it easily falls to a predetermined position at the bottom of the container, and a liquid sample is introduced. At this time, since the sample adhering to the wall surface is likely to fall off, it is preferable that the cylindrical body portion and the tapered detail having a conical shape are combined.

[Reagent holder]
In the tube container, the reagent or a carrier carrying the reagent is dried or frozen in the tapered portion so that the reagent can remain in the vicinity of a predetermined reaction zone or detection zone of the tube vessel. A reagent holding part that can be held in that position by drying is provided integrally with the reaction tube.
It is desirable that the detailed installation location of the reagent holding unit is appropriately adjusted according to the internal structure of the reaction / detection device to be used.

Any reagent holding unit may be used as long as the reagent holding unit has the above-described function as long as the space of the reaction tube is not vertically divided by the reagent holding unit (that is, the two spaces do not communicate above and below the reagent holding unit). It may be formed in any shape.
For example, the reagent holding part is formed so as to project from the inner wall of the tube container toward the inside of the tube container, but the part is formed without reaching the opposing wall surface ("extended part" described later). , When formed to reach the opposite wall surface (for example, when the reagent holding part is formed in a cross shape when viewed from the top of the tube container), or a recess is formed from the inner wall of the container toward the inner wall. And so on (“recessed part” to be described later).

As one form of the reagent holding part, there is an “overhanging part” that protrudes from the inner wall surface of the tapered portion toward the center of the reaction tube, but does not reach the opposing wall surface.
It is desirable that 2 to 10 and more preferably 3 to 6 overhang portions are disposed to face each other over the inner periphery of the tapered portion.
The overhanging portion may extend from the lower portion of the tapered portion to the bottom portion, or may not be extended to the bottom portion and may be formed in a rib shape. When the overhanging portion extends to the bottom, the side surface of the overhanging portion on the tube center side may be extended in parallel with the longitudinal center line of the reaction tube, or inclined toward the bottom. (For example, inclined downward toward the longitudinal center line of the reaction tube).

  When viewed from the cross section of the reaction tube, the shape of the projecting portion is, for example, a strip shape in which a rectangle is elongated, a triangular shape in which a tip projecting toward the inside of the container is sharp, and a table having a flat tip The shape may be various, such as a dome shape with a rounded tip, or a concave shape with a tip narrowed toward the inner wall of the container. Further, the overhanging part may be formed to have a skirt-like shape from the aforementioned tip toward the inner wall surface of the container.

  Moreover, although the upper surface of the said overhang | projection part may be a horizontal plane, it may be formed inclining downward toward the center of a tube.

Another embodiment of the reagent holding portion includes a “concave portion” formed on the inner wall of the tapered portion from the inner wall toward the inside of the inner wall over the inner periphery of the tapered portion.
It is desirable that 2 to 10 recesses, more preferably 3 to 6 are disposed so as to face each other over the inner periphery of the tapered portion.
Moreover, it is desirable that the concave portion extends from the lower portion of the tapered portion to the bottom portion. At this time, the side surface of the recess on the tube center side may be extended in parallel with the center line in the longitudinal direction of the reaction tube, or may have an inclination toward the bottom (for example, the center in the longitudinal direction of the reaction tube). It may be extended (inclined downwards towards the line).

  When viewed from the cross section of the reaction tube, the shape of the concave portion is, for example, a strip shape in which a rectangle is elongated, a triangular shape in which a tip protruding toward the inside of the container is sharp, and a trapezoid shape in which the tip is flat The shape may be various, such as a dome shape having a rounded tip, or a concave shape having a tip narrowed toward the inner wall of the container. Further, the recess may be formed to have a skirt-like shape from the aforementioned tip toward the inner wall surface of the container.

  Moreover, although the upper surface of the said recessed part may be a horizontal plane, it may be formed inclining downward toward the center of a tube.

  The shape and size of the overhang / recess in the reagent holding unit described above, and the number of installed portions are not only easy to manufacture the tube container, but also easy to adhere the dry reagent, and the reagent is dried and solidified in the container. It is determined taking into account various factors such as the ease of entering the uneven shape and the ease of being caught.

[Reagent kit]
The present invention also relates to a reagent kit comprising the aforementioned tube container and a dry reagent held in the reagent holding part of the container or a carrier capable of carrying the dry reagent.

As described above, the dry reagent included in the reagent kit is a carrier that carries a reagent by loading a liquid reagent toward a support such as a nonwoven fabric adhered to the reagent holding unit, and the carrier is used under reduced pressure. Examples thereof include a dried product and a product obtained by putting a liquid reagent into a tube container and drying it in the container.
The above reagent is preferably a nucleic acid amplification reagent. By using an antigen, antibody, dye, enzyme, substrate, etc., the reagent kit of the present invention can be used for measuring an immunoreaction or biochemical reaction, by immunofluorescence. It can also be used for trace detection.

Examples of the amplification method that can be performed with the nucleic acid amplification reagent include LAMP, PCR, NASBA, LCR, SDA, RCA, ICAN method and the like.
When a nucleic acid amplification reagent is employed as the reagent, an amplified nucleic acid detection reagent may further be included as desired. Examples of detection methods that can be carried out by the amplified nucleic acid detection reagent include a fluorescence method, a fluorescence polarization method, a turbidity method, a colorimetric method, and a luminescence method.

  The carrier capable of carrying the dry reagent is formed by applying or immersing the above-described reagent on a support made of filter paper, membrane filter, paper, thread, nonwoven fabric, or the like, and then drying under reduced pressure, for example. The size is desirably a size that can be positioned on the reagent holding portion of the tube container (a size that does not fall below the reagent holding portion).

In the reagent kit of the present invention, the tube container holding the dry reagent (or the carrier capable of supporting the dry reagent) may be formed with only one tube, or a plurality of tube containers may have a connecting portion. You may have the structure arranged in a line through.
The connecting portion is formed so as to connect a plurality of lid portions, joint portions, reaction tubes, or a combination thereof, but is preferably formed so as to connect a plurality of reaction tubes. It is desirable. At this time, the connecting portion may be formed so as to be connected at an edge provided at the opening of the reaction tube, or may be formed so as to be connected at a cylindrical body portion of the reaction tube.

As an example, a case where the reagent kit of the present invention is used for nucleic acid amplification / detection reaction is shown below.
In the reagent kit, the sample for detection is charged toward the dried reagent held in the reagent holding unit, and then the reagent and the container into which the sample has been introduced are heated to perform nucleic acid amplification and amplification. Detect nucleic acids.

  Here, after putting the sample toward the dry reagent (carrier holding the dry reagent, 50), the schematic diagram of the inside of the tube container after stirring by the stirrer is shown in FIG. 12b (conventional tube container) and FIG. The tube container of the present invention). As can be seen from these figures, when a conventional tube container having no reagent holding part is used (FIG. 12b), after sample input, stirring, etc., various parts of the inner wall inside the container are placed. The dry reagent brought into contact with the sample adheres, but when the tube container of the present invention is used, the reagent stays below the container (reagent holding part). For this reason, the tube container of the present invention not only saves the trouble of pushing and dropping the dry reagent brought into contact with the sample into a predetermined reaction zone / detection zone (lower part of the vessel) but also adheres to the inner wall of the tube vessel. It is difficult to cause measurement failure and accuracy failure caused by this.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated in more detail, these are examples and this invention is not limited to these Examples at all.

<Example 1>
FIG. 1 is a plan view (FIG. 1 a) viewed from above the tube container 1 of the present invention and a longitudinal sectional view cut along the line XX ′ in FIG. 1 a (FIG. 1 b, hereinafter “vertical sectional view”). Indicates a similar cut surface). In a preferred embodiment of the tube container 1 of the present invention, the reaction tube 10 is integrally molded so as to be coupled to the lid portion 30 having the display portion 31, the seal portion 32, and the outer peripheral portion 33 via the joint portion 40. Here, the thickness of the joint portion 40 is reduced at the approximate center so that it can be easily bent.
The reaction tube 10 is provided with a cylindrical body 11 continuous from the opening 14, a taper 12 formed to be tapered from the body 11, and a bottom 13 formed so as to close the tip of the taper. ing. In the opening portion 14, an edge portion 16 that can further enhance the adhesion of the seal portion 32 by being in close contact with the outer peripheral portion 33 of the lid portion 30 is formed. The tapered portion 12 is provided with a reagent holding unit 20 so as to be integrated.

<Example 2>
FIG.2 and FIG.3 is a figure showing the longitudinal cross-sectional view of this invention tube container 1 provided with the reagent holding | maintenance part 20 of a 1st form.
2A to 2E, the reagent holding unit 20 includes a protruding portion 21 (a to e) that protrudes from below the tapered portion 12 toward the center (center line 2) of the reaction tube 10.
2a, 2c and 2e, the upper surfaces 212a, 212c and 212e of the overhang portions 21a, 21c and 21e are formed in a horizontal plane. Alternatively, as shown in FIGS. 2 b and 2 d, the upper surfaces 212 b and 212 d of the overhang portions 21 b and 21 d may be formed to be inclined downward from the inner wall 15 of the reaction tube 10 toward the center line 2.
Further, as shown in FIG. 2e, the side surface 211e on the center side of the reaction tube 10 of the overhang portion 21e may be formed to be inclined downward toward the center line 2.
Further, the overhanging portion 21 extends from the bottom of the tapered portion 12 to the bottom portion 13 (see FIGS. 2a, 2b and 2e), but does not extend to the bottom portion 13 (FIGS. 2c and 2d). However, it is preferable to extend to the bottom 13 as shown in FIGS. 2a, 2b and 2e.

  3A and 3B, the reagent holding part 20 is formed as rib-like protruding parts 22a and 22b. The shape of the rib-like overhanging portions 22a and 22b may be either a protrusion (22a) or a plate-like protrusion (22b) whose width changes toward the bottom 13 of the reaction tube 10.

<Example 3>
The top views seen from the upper side of the tube container 1 provided with the reagent holding part 20 of said 1st form are shown in FIGS.
As illustrated in FIGS. 4A to 4G, the overhang portion 21 or the rib-like overhang portion 22 provided as the reagent holding portion 20 is provided, for example, at 2 to 8 locations on the inner wall 15 of the reaction tube 10. . Considering the ease of molding of the reaction tube 10 and the stability of the retained dry reagent, most preferably 3 to 6 (FIGS. 4b to 4e) overhangs 21 or over the inner periphery of the inner wall 15 It is preferable that the rib-like overhang portions 22 are arranged so as to face each other.

Further, as illustrated in FIGS. 5A to 5F, when the overhanging portion 21 or the rib-like overhanging portion 22 is viewed from above, the shape thereof is a rectangular shape (FIG. 5a), and the rectangular shape with a rounded tip (FIG. 5b). , Triangle (FIG. 5c), trapezoid (FIG. 5d), dome shape (FIG. 5e), concave shape (FIG. 5f), etc.
Furthermore, as illustrated in FIGS. 6A to 6C, when the overhang portion 21 or the rib-like overhang portion 22 is viewed from above, the thickness (FIG. 6 a) and the length (FIG. 6b) may be different, or a combination of the shapes shown in FIGS. 5a to 5f, for example, a shape combining triangles and squares (FIG. 6c).

<Example 4>
7A to 7C are a plan view (FIGS. 7a and 7b) and a vertical cross-sectional view (FIGS. 7c and 7a) of the tube container 1 of the present invention provided with the reagent holding unit 20 of the second form. (Corresponding to a longitudinal sectional view), and the reagent holder 20 is provided with a recess 23.
The number of the recessed portions 23 may be an even number (for example, four, FIG. 7a) or an odd number (for example, three, FIG. 7b). Similar to the overhang portion 21 described above, considering the ease of molding of the reaction tube 10 and the stability of the retained dry reagent, it is most preferable that the three are arranged so as to face each other over the inner circumference of the inner wall 15 (see FIG. 7b), 4 (FIG. 7a), 5 and 6 are preferably provided.
Moreover, the shape of the recessed part 23 may be trapezoidal as shown in FIG. 7a, and the shape may be various, such as a strip shape, a triangle, and a dome shape, like the above-described overhanging portion 21.
Further, as shown in FIG. 7 c of the longitudinal cross section of the tube container 1 of the present invention having the reagent holding unit 20 of the second form, the concave portion 23 extends so as to reach the bottom portion 13 from below the tapered portion 12 of the reaction tube 10. However, it may not be installed so as to reach the bottom portion 13.

<Example 5>
FIGS. 8A to 8D are plan views of the lid 30 that seals the opening 14 of the reaction tube 10 together with the joint 40 and the opening 14 as viewed from above.
As shown in FIGS. 8a to 8d, the lid portion 30 is provided with a display portion 31 on which a sign indicating the contents of the reaction tube 10 can be written. Since the display unit 31 has a relatively large area, the display unit 31 can be easily pinched with a finger, and can easily be attached to and detached from the opening 14 of the seal unit 32, and thus also serves as a tab.
The shape of the display unit 31 is not particularly limited. For example, as shown in FIG. 8, the display unit 31 may have a shape such as a substantially rectangular shape (FIGS. 8 a and 8 c) or a substantially triangular shape (FIGS. 8 b and 8 d). Further, the direction of the display unit 31 may be the same direction (FIGS. 8a and 8b) as that of the bonding unit 40 or a different direction (FIGs. 8c and 8d).
Preferably, it is easy to attach a reference numeral 311 for identifying the sample to the display unit 31 (see FIG. 10 to be described later). It is desirable to use a substantially rectangular shape.

<Example 6>
An example of the reagent kit of the present invention is shown in FIG. 9 (front view) and FIG. 10 (perspective view from above).
As shown in FIGS. 9 and 10, the reagent kit 70 includes a tube container 1 and a carrier 50 holding a dry reagent held in a reagent holding unit 20 inside a reaction tube 10 of the tube container.
As shown in FIGS. 9 and 10, the reagent kit 70 is configured as a series of reagent kits 70 in which the upper portion of the tube container 1 is arranged in a horizontal row via the connecting portion 60. In addition, the length of the connection part 60 shown in FIG.9 and FIG.10 and the number of the containers 1 contained in a series of reagent kits change suitably according to the apparatus to be used.

<Use Example 1>
FIG. 11 is a view showing a dry reagent 51 that is solidified by putting a reagent into the tube container 1 of the present invention and freeze-drying in the container, and shows a longitudinal sectional view of the tube container 1. By lyophilization, the volume of the reagent (liquid) slightly increases, and the dry reagent 51 is solidified so as to be sandwiched between the reagent holding portions 20.

<Usage example 2>
Nucleic acid was amplified by the LAMP method using the tube container of the present invention.
[Reaction solution]
The basic composition of the reaction solution for amplifying nucleic acids by the LAMP method and the primer sequences of the reagents are shown in the table below.

[Nucleic acid amplification reaction example 1: change in carrier of dry reagent]
A non-woven fabric having a diameter of 3 mm (material: rayon (Ry) or polyvinyl alcohol (PVA)) was employed as the carrier, and after applying the dry reagent to each carrier, drying under reduced pressure was performed to prepare a reduced-pressure dry reagent. A liquid reagent is added to each of these dry reagents to form a LAMP reaction solution, and MTB plasmid DNA added at 100 copies / tube is “with target gene”, and those without added are “no target gene”. The LAMP reaction was performed at 67 ° C. for 60 minutes.
Using rayon nonwoven fabric as a support, the result of LAMP reaction (measurement device: using real-time fluorescence measurement device FluoDia T70 (manufactured by Otsuka Electronics Co., Ltd.)) is shown in FIG. FIG. 14 shows the results of the LAMP reaction (measurement apparatus: using a real-time turbidity measurement apparatus LA-320C (manufactured by Eiken Chemical Co., Ltd.)). In addition, in order to dissolve a polyvinyl alcohol nonwoven fabric, the measurement was not performed for 5 minutes at the beginning of the reaction.
According to this, even when a vacuum dried reagent using a rayon nonwoven fabric (insoluble nonwoven fabric) as a reagent carrier is used (FIG. 13), a vacuum dried reagent using a polyvinyl alcohol nonwoven fabric (soluble nonwoven fabric) as a reagent carrier is used. In any of (FIG. 14), it was possible to detect the target gene.

[Nucleic acid amplification reaction example 2: change in shape of reagent holding unit installed in reaction tube]
The above-mentioned dry reagent was dispensed into reaction tubes each having a rib (see FIG. 3a) or an overhanging portion (see FIG. 2a) as a reagent holding part inside, and then dried under reduced pressure to prepare a vacuum dry reagent. A liquid reagent is added to this dry reagent to give a LAMP reaction solution, and MTB plasmid DNA added so as to be 100 copies / tube is “with target gene”, and those without added are “no target gene”, Using a real-time turbidity measuring device RT-160C (manufactured by Eiken Chemical Co., Ltd.), LAMP reaction was carried out at 67 ° C. for 60 minutes.
As a result, as shown in FIG. 15 (rib) and FIG. 16 (overhang portion), it was possible to detect the target gene even when the reagent holding portion had any shape.

The base sequence of the primer used in this example is shown in the following SEQ ID No. in the sequence listing.
SEQ ID NO: 1 Mycobacterium tuberculosis H37Rv Gyr B sequence (GenBank: BX842572)
SEQ ID NO: 2 F1
SEQ ID NO: 3 F2
SEQ ID NO: 4 F3
SEQ ID NO: 5 Loop F (LPF)
Sequence number 6 B1c
Sequence number 7 B2c
Sequence number 8 B3c
SEQ ID NO: 9 Loop B (LPB)
SEQ ID NO: 10 FIP
Sequence number 11 BIP
SEQ ID NO: 12 B3

1 (a, b) is a plan view (FIG. 1a) viewed from above the tube container 1 of the present invention and a longitudinal sectional view (FIG. 1b) cut in the longitudinal direction along the line XX ′ of FIG. 1a. FIG. Fig.2 (ae) is a figure which shows the longitudinal cross-sectional view of the tube container 1 of this invention provided with the reagent holding | maintenance part 20 (overhang | projection part 21) of a 1st form. FIG. 3 (a, b) is a view showing a longitudinal sectional view of the tube container 1 of the present invention including the reagent holding part 20 (rib-shaped overhanging part 22) of the first form. FIGS. 4A to 4G are views showing a plan view of the tube container of the present invention provided with the reagent holding unit 20 of the first form as viewed from above. FIGS. 5A to 5F are views showing a plan view of the tube container of the present invention provided with the reagent holding unit 20 of the first form as seen from above. 6A to 6C are plan views of the tube container 1 of the present invention including the reagent holding unit 20 according to the first embodiment as viewed from above. 7A to 7C are a plan view (FIGS. 7a and 7b) and a longitudinal section (FIG. 7c) as viewed from above the tube container 1 of the present invention including the reagent holding unit 20 of the second form. 8A to 8D are plan views of the lid 30 as viewed from above together with the joint 40 and the opening 14. FIG. 9 is a front view of the reagent kit 70 of the present invention. FIG. 10 is a perspective view of the reagent kit 70 of the present invention as viewed from above. FIG. 11 is a longitudinal sectional view of the tube container 1 in which the dry reagent 51 solidified by freeze-drying in the container is held in the reagent holding unit 20. FIG. 12 is a front view showing the presence or absence of the rising of the dry reagent in comparison between the tube container of the present invention (FIG. 12a) and the conventional tube container (FIG. 12b). FIG. 13 shows measurement results of nucleic acid amplification by fluorescence measurement of a carrier carrying a dry reagent with an insoluble nonwoven fabric (rayon) as a support. FIG. 14 shows the results of nucleic acid amplification measurement by turbidity measurement of a carrier carrying a dry reagent with a soluble nonwoven fabric (polyvinyl alcohol) as a support. FIG. 15 shows a result of nucleic acid amplification by turbidity measurement performed using a tube having a rib as a reagent holding unit. FIG. 16 shows the result of nucleic acid amplification by turbidity measurement performed using a tube having an overhanging portion as a reagent holding portion.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Tube container 2 ... Longitudinal centerline 10 of a tube container ... Reaction tube,
11 ... trunk,
12 ... Fine details,
13 ... bottom,
14 ... opening,
15 ... inner wall,
16 ... edge,
20 ... Reagent holding part,
21 (ae) ... overhang part,
211 (ae) ... the side surface of the overhanging portion on the center side of the reaction tube,
212 (ae) ... the upper surface of the overhang,
22 (a, b) ... rib-like overhanging part,
23 ... concave portion,
30 ... lid,
31 ... display part,
311... Code for identifying the sample,
32 ... seal part,
33 ... outer periphery,
40 ... Junction part,
50 ... a carrier holding a dry reagent,
51 ... A dry reagent solidified by freeze-drying in a container,
60 ... connecting part,
70: Reagent kit.

Claims (12)

A tube container for reaction or detection of a target substance in a small amount of sample,
A reaction tube, a lid portion that seals the opening on the upper surface thereof, and a joint portion that joins the upper portion of the reaction tube and the lid portion are integrally formed,
The reaction tube has a cylindrical body part that is continuous from the opening part, a tapered part that is tapered from the trunk part, and a bottom part that is formed so as to close a tip of the tapered part, A tube container in which a reagent holding part capable of holding a reagent or a carrier carrying a reagent in the inside of the tapered portion is held at that position by drying or freeze-drying of the reagent. 2. The tube container according to claim 1, wherein the reagent holding portion is a protruding portion that protrudes from the inner wall surface of the tapered portion toward the center of the reaction tube. 3. The tube container according to claim 2, wherein 2 to 10 overhang portions are provided to face each other over an inner periphery of the tapered portion. 4. The tube container according to claim 3, wherein three to six of the overhang portions are provided to face each other over an inner periphery of the tapered portion. The tube container according to any one of claims 2 to 4, wherein the projecting portion extends from a lower portion of the tapered portion to the bottom portion. The tube container according to any one of claims 2 to 4, wherein the overhang portion is formed of a rib. The tube container according to claim 1, wherein the reagent holding portion is formed of a concave portion formed on an inner wall of the tapered portion over an inner periphery of the tapered portion. The tube container according to claim 7, wherein 2 to 10 concave portions are provided to face each other over the inner periphery of the tapered portion. The tube container according to claim 8, wherein 3 to 6 concave portions are provided to face each other over the inner periphery of the tapered portion. The tube container according to any one of claims 7 to 9, wherein the concave portion extends from a lower portion of the tapered portion to the bottom portion. The tube container according to any one of claims 1 to 10, wherein a display area having a larger area for indicating the contents of the container is provided on the lid. A reagent kit comprising the tube container according to any one of claims 1 to 11 and a dry reagent or a carrier capable of supporting the dry reagent held in a reagent holding part of the tube container.