JP2017201932A - Reagent cartridge for analysis of target nucleic acid, apparatus for analyzing target nucleic acid, and method for analyzing target nucleic acid - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a reagent cartridge capable of analyzing at a component concentration suitable for oligonucleotide for analysis of each target nucleic acid, in analysis of a plurality of target nucleic acids, to provide an apparatus for analyzing a target nucleic acid, and to provide a method for analyzing a target nucleic acid.SOLUTION: A reagent cartridge for analyzing a target nucleic acid is disclosed, a cartridge 1 for analyzing a first target nucleic acid having at least one common reagent bath 11, a plurality of concentration adjustment reagent baths 12, and a plurality of non-common reagent baths 13, the common reagent bath(s) 11 being filled with a common reagent respectively containing a plurality of components usable in common with analysis of a plurality of target nucleic acids, the plurality of concentration adjustment reagent baths 12 being filled with a concentration adjustment reagent respectively containing at a different concentration at least one component usable in common with analysis of a plurality of target nucleic acids, and the plurality of non-common reagent baths 13 being capable of being filled with a non-common reagent containing oligonucleotide for analysis for analyzing a particular target nucleic acid.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a target nucleic acid analysis reagent cartridge, a target nucleic acid analysis apparatus, and a target nucleic acid analysis method.

  Patent Document 1 includes a common reagent tank filled with a common reagent that can be commonly used in gene analysis such as PCR, and a non-analysis oligonucleotide such as a primer prepared by a user according to a target nucleic acid. A reagent cartridge having a non-common reagent tank capable of being filled with a common reagent is described.

Patent No. 5498944

  In Patent Document 1, one common reagent tank is provided for each reagent, and each tank is filled with a reagent of a predetermined concentration. For this reason, if it is necessary for a certain component to have different concentrations even with the same reagent in accordance with the characteristics of the primer, probe, etc., for example, if a component having a concentration lower than the predetermined concentration is required, In the cartridge of Document 1, since the concentration of the component is a predetermined concentration, it is difficult to make only the component have a different concentration.

  Furthermore, when measuring a plurality of target nucleic acids such as a plurality of target genes for one sample, an analysis system having different component concentrations corresponding to each target nucleic acid analysis oligonucleotide is prepared in one cartridge. However, it is difficult to cope with the cartridge of Patent Document 1.

  Therefore, the present invention provides a reagent cartridge, a target nucleic acid analyzer, and a target nucleic acid analysis method capable of performing analysis at a component concentration suitable for an oligonucleotide for analysis of each target nucleic acid in analysis of a plurality of target nucleic acids. The purpose is to provide.

In order to solve the problems of the present invention, a reagent cartridge for analyzing a first target nucleic acid (hereinafter also referred to as “cartridge”) of the present invention comprises at least one common reagent tank, a plurality of concentration adjusting reagent tanks, A plurality of non-common reagent vessels,
Each of the common reagent tanks is filled with a common reagent containing a plurality of components that can be used in common for analysis of a plurality of target nucleic acids,
Each of the plurality of concentration adjustment reagent tanks is filled with a concentration adjustment reagent containing at least one component that can be commonly used for analysis of a plurality of target nucleic acids at different concentrations,
Each of the plurality of non-common reagent tanks can be filled with a non-common reagent containing an analytical oligonucleotide for analyzing a specific target nucleic acid.

The second cartridge of the present invention has a plurality of common reagent reservoirs and a plurality of non-common reagent reservoirs,
Each of the common reagent tanks is filled with a common reagent containing a plurality of components that can be used in common for analysis of a plurality of target nucleic acids,
Each of the common reagents includes at least one component of the plurality of components at different concentrations,
Each of the plurality of non-common reagent tanks can be filled with a non-common reagent containing an analytical oligonucleotide for analyzing a specific target nucleic acid.

A first target nucleic acid analyzer of the present invention (hereinafter, also referred to as “analyzer”) includes a cartridge housing portion in which the first cartridge of the present invention is housed,
Mixing a sample, a common reagent filled in the common reagent tank, at least one concentration adjusting reagent filled in the concentration adjusting reagent tank, and at least one non-common reagent filled in the non-common reagent tank A preparation unit for preparing an analysis system for the target nucleic acid,
And an analysis unit for analyzing the target nucleic acid by analyzing the analysis system.

The first target nucleic acid analysis method of the present invention (hereinafter also referred to as “analysis method”) uses the first cartridge of the present invention,
Mixing a sample, a common reagent filled in the common reagent tank, at least one concentration adjusting reagent filled in the concentration adjusting reagent tank, and at least one non-common reagent filled in the non-common reagent tank A preparation step of preparing an analysis system for the target nucleic acid,
An analysis step of analyzing the target nucleic acid by analyzing the analysis system.

  According to the present invention, in analysis of a plurality of target nucleic acids, a reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Therefore, according to the present invention, for example, the target nucleic acid can be analyzed with better analysis accuracy.

FIG. 1A is a perspective view illustrating a configuration of a cartridge according to the first embodiment, and FIG. 1B is a cross-sectional view as viewed from the II direction of FIG. FIG. 2A is a perspective view showing the configuration of the cartridge of Embodiment 2, and FIG. 2B is a cross-sectional view as seen from the II-II direction of FIG. FIG. 3A is a perspective view showing the configuration of the cartridge according to the third embodiment, and FIG. 3B is a cross-sectional view taken from the III-III direction of FIG. FIG. 4 is a block diagram illustrating a configuration of the analyzer according to the fourth embodiment.

In the first cartridge of the present invention, for example, the components contained at different concentrations in the concentration adjusting reagent are:
In the analysis of the target nucleic acid, it is a component whose concentration is adjusted according to the type of analysis oligonucleotide of the non-common reagent used in combination with the common reagent and the concentration adjusting reagent.

  In the first cartridge of the present invention, for example, the common reagent does not include a component that is contained at a different concentration in the concentration adjusting reagent.

In the second cartridge of the present invention, for example, the components contained in the common reagent at different concentrations are
In the analysis of the target nucleic acid, it is a component whose concentration is adjusted according to the kind of the oligonucleotide for analysis of the non-common reagent used in combination with the common reagent.

  In the first and second cartridges of the present invention, for example, the components contained in the different concentrations are at least one selected from the group consisting of magnesium ions, potassium ions, manganese ions, sodium ions, and ammonium ions. Preferably, it is a magnesium ion.

  In the first and second cartridges of the present invention, for example, the oligonucleotide for analysis is a primer capable of amplifying the target nucleic acid, and preferably further includes a probe capable of detecting the target nucleic acid.

  In the first and second cartridges of the present invention, for example, the non-common reagent tank is pre-filled with the non-common reagent.

  In the first and second cartridges of the present invention, for example, each non-common reagent filled in each non-common reagent tank contains oligonucleotides for analyzing different target nucleic acids.

  The first analyzer of the present invention prepares the analysis system by, for example, mixing the sample, at least two concentration adjusting reagents, and at least one non-common reagent in the preparation unit.

  In the analysis method of the present invention, for example, in the preparation step, the analysis system is prepared by mixing the sample, at least two concentration adjusting reagents, and at least one non-common reagent.

<First reagent kit for analyzing target nucleic acid>
As described above, the first cartridge of the present invention has at least one common reagent tank, a plurality of concentration-adjusting reagent tanks, and a plurality of non-common reagent tanks. Are filled with a common reagent containing a plurality of components that can be commonly used for analysis of target nucleic acids, and each of the plurality of concentration adjusting reagent tanks is at least commonly used for analysis of a plurality of target nucleic acids. Concentration adjustment reagents containing different concentrations of one component are filled, and each of the plurality of non-common reagent tanks can be filled with a non-common reagent containing an analysis oligonucleotide for analyzing a specific target nucleic acid. It is characterized by. The first cartridge of the present invention has a plurality of concentration adjustment reagent tanks, and each of the plurality of concentration adjustment reagent tanks has at least one component that can be commonly used for analysis of a plurality of target nucleic acids at different concentrations. It is characterized by being filled with a concentration adjusting reagent contained in the above, and other configurations and conditions are not particularly limited.

  As described above, the first cartridge of the present invention has a plurality of concentration adjustment reagent tanks, and each of the plurality of concentration adjustment reagent tanks can be used in common for analysis of a plurality of target nucleic acids. A concentration-adjusting reagent containing two components at different concentrations is packed. For this reason, for example, when analyzing two target nucleic acids, the components contained in the different concentrations (hereinafter also referred to as “adjusting components”) are optimal according to the type of the oligonucleotide used for analyzing each target nucleic acid. By setting the appropriate concentration, the sample, the common reagent, the concentration adjusting reagent set to the optimum concentration, and the non-common reagent containing the corresponding analytical oligonucleotide are mixed in a predetermined amount. Thus, an analysis system suitable for the analysis oligonucleotide can be easily prepared. Also, when analyzing three or more target nucleic acids, one of the adjustment components is set to the highest component concentration or higher among the optimum concentrations according to the type of oligonucleotide for analysis for analyzing each target nucleic acid. However, by setting another one below the lowest component concentration among the optimal concentrations according to the type of oligonucleotide for analysis for analyzing each target nucleic acid, for example, these two adjustment components An analysis system suitable for the oligonucleotide for analysis can be easily prepared by mixing a concentration adjusting reagent containing a sample, the common reagent, and the non-common reagent in a predetermined amount. Therefore, according to the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Furthermore, according to the first cartridge of the present invention, since the adjustment component can be an analysis system suitable for the analysis oligonucleotide, the target nucleic acid can be analyzed with better analysis accuracy, for example.

  In the present invention, the analysis may be, for example, a qualitative analysis for determining the presence or absence of the target nucleic acid, or a quantitative analysis for determining the amount of the target nucleic acid.

  In the present invention, the material of the cartridge is not particularly limited, and examples thereof include plastic.

  In the present invention, the common reagent includes a plurality of components that can be commonly used for analysis of a plurality of target nucleic acids. The plurality of components contained in the common reagent are not particularly limited, and can be appropriately determined according to, for example, the target nucleic acid analysis method. When the analysis method includes a nucleic acid amplification method such as PCR, the plurality of components included in the common reagent include, for example, reagents used in the nucleic acid amplification method. Specific examples include a DNA polymerase such as a heat-resistant DNA polymerase, Examples include deoxynucleotide triphosphate (dNTP), albumin such as bovine serum albumin, and preservatives such as sodium azide. The concentration of the plurality of components contained in the common reagent is not particularly limited, and can be appropriately set according to, for example, each analysis method. It is preferable that the common reagent does not contain components that are contained at different concentrations in the concentration adjusting reagent described later. This makes it possible to more easily adjust the concentration of the adjustment component in the analysis system in which the sample, the common reagent, the concentration adjusting reagent, and the non-common reagent are mixed. The common reagent may be, for example, a liquid reagent or a solid reagent (dry reagent). In the former case, the common reagent is, for example, a reagent in which the common reagent is dissolved, suspended, or mixed in a solvent. The solvent is not particularly limited, and examples thereof include buffers such as water, Tris-HCl, Tricine, MES, MOPS, HEPES, and CAPS. Specific examples include commercially available PCR buffers and commercially available PCR kits. And the like.

  In the present invention, the target nucleic acid may be, for example, DNA or RNA. The analysis of the target nucleic acid can also be referred to as analysis of an analysis target in the target nucleic acid, for example. The analysis target in the target nucleic acid is not particularly limited, and examples thereof include single nucleotide polymorphism (SNP), gene, gene mutation, non-coding RNA, chromosome, chromosome abnormality and the like. Examples of the gene mutation include deletion, substitution, insertion, and / or addition of one or more bases in the gene. Examples of the chromosomal abnormality include chromosome fragmentation, deletion, duplication, inversion, translocation, translocation, attachment, and / or deletion. In the present invention, the target nucleic acid may be, for example, one or plural. In the former case, the target nucleic acid includes, for example, a plurality of analysis objects.

  In the present invention, the concentration adjusting reagent contains at least one component that can be commonly used for analysis of a plurality of target nucleic acids at different concentrations. The adjustment component can be said to be a component whose concentration is adjusted according to the type of oligonucleotide for analysis of the non-common reagent used in combination with the common reagent and the concentration adjustment reagent in the analysis of the target nucleic acid, for example. . The concentration of the adjustment component in the concentration adjustment reagent can be appropriately set by those skilled in the art, for example, according to the base sequence of the oligonucleotide for analysis. In addition, the concentration ratio of the adjustment component between the concentration adjustment reagents is not particularly limited, and can be appropriately set according to, for example, the analysis oligonucleotide. Specific examples of the adjustment component include magnesium ion, potassium ion, manganese ion, sodium ion, ammonium ion, and the like, and preferably magnesium ion. In the concentration adjusting reagent, the adjusting component is, for example, an ion derived from a salt contained in the concentration adjusting reagent. The adjustment component may be one or plural. The number of the concentration adjusting reagents may be plural, for example, 2 to 4. Each of the plurality of concentration adjusting reagents may include at least one component at a different concentration, and the concentrations of the other components may be the same or different. When the adjustment component is magnesium ion, the concentration of magnesium ion in each concentration adjustment reagent is, for example, a concentration at which the concentration in the analysis system is 1 to 10 mmol / L. When the adjustment component is potassium ion, the concentration of potassium ion in each concentration adjustment reagent is, for example, such that the concentration in the analysis system is 0 to 75 mmol / L, exceeds 0 mmol / L, and is 75 mmol / L or less. When the adjustment component is sodium ion, the concentration of sodium ion in each concentration adjustment reagent is, for example, such that the concentration in the analysis system is 0 to 75 mmol / L, more than 0 mmol / L, and 75 mmol / L or less. When the adjustment component is manganese ions, the concentration of manganese ions in each concentration adjustment reagent is, for example, such that the concentration in the analysis system is 0 to 1 mmol / L, more than 0 mmol / L, and 1 mmol / L or less. When the adjustment component is ammonium ions, the concentration of ammonium ions in each concentration adjustment reagent is, for example, a concentration at which the concentration in the analysis system is 5 to 30 mmol / L. When the number of the concentration adjusting reagents is 3 or more, the concentration of the adjusting component in any two concentration adjusting reagents may be the same. The concentration adjusting reagent may be, for example, a liquid reagent or a solid reagent (dry reagent). In the former case, the concentration adjusting reagent is, for example, a reagent obtained by dissolving, suspending, or mixing the concentration adjusting reagent in a solvent. As the solvent, for example, the above description can be used. For example, the solvent may be the same as or different from the common reagent. When the concentration adjusting reagent is a solid reagent, the concentration of the adjusting component is, for example, the concentration when dissolved, suspended, or mixed in a predetermined amount of solvent in the cartridge.

  In the present invention, the non-common reagent includes the analysis oligonucleotide for analyzing the specific target nucleic acid. In the present invention, the analytical oligonucleotide is not particularly limited, and can be appropriately determined according to, for example, the type of the target nucleic acid. As a specific example, the oligonucleotide for analysis includes, for example, a primer. The primer may be, for example, a primer capable of amplifying the target nucleic acid or a primer capable of amplifying a polynucleotide containing the target nucleic acid. As a specific example, a forward primer and a reverse primer capable of amplifying the target nucleic acid And a primer set including a forward primer and a reverse primer capable of amplifying a polynucleotide including the target nucleic acid. When the analysis oligonucleotide includes the primer, the analysis oligonucleotide may further include a probe in addition to or instead of the primer. The probe is, for example, a probe that can detect the target nucleic acid, and more specifically, a probe that can detect the target nucleic acid in an amplified fragment amplified by the primer. When the analytical oligonucleotide includes the primer and probe, the primer and probe may be placed in separate non-common reagent vessels. The primer and the probe may be labeled with a labeling substance, for example. In the present invention, the base sequences of the primer and the probe are not particularly limited, and can be appropriately designed by those skilled in the art according to, for example, the type of the target nucleic acid and the base sequence thereof.

  The non-common reagent may include, for example, a part of a plurality of components that can be commonly used for analysis of the plurality of target nucleic acids included in the common reagent. As a specific example, the analysis method is a nucleic acid amplification method. The non-common reagent may contain dNTPs. In this case, it is preferable that the common reagent does not contain dNTP. The non-common reagent may be, for example, a liquid reagent or a solid reagent (dry reagent). In the former case, the non-common reagent is, for example, a reagent obtained by dissolving, suspending, or mixing the non-common reagent in a solvent. As the solvent, for example, the above description can be used. For example, the solvent may be the same as or different from the common reagent and / or the concentration adjusting reagent.

  Hereinafter, a first cartridge of the present invention and an analysis method using the same will be described in detail with reference to the drawings. However, the present invention is not limited or limited to the following examples. In the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown, unlike the actual case. Each embodiment can be combined with each other unless otherwise specified.

(Embodiment 1)
1A and 1B are schematic views of a cartridge according to a first embodiment. FIG. 1A is a perspective view of the cartridge according to the first embodiment, and FIG. 1B is a view in the II direction of FIG. It is sectional drawing. As shown in FIG. 1A, the cartridge 1 of the present embodiment includes a common reagent tank 11, a concentration adjusting reagent tank 12, and a non-common reagent tank 13. Further, as shown in FIG. 1B, the common reagent tank 11 is filled with a common reagent 111 containing a plurality of components that can be used in common for the analysis of the plurality of target nucleic acids. The concentration adjustment reagent tank 12 is divided into concentration adjustment reagent tanks 12a and 12b, and at least one component that can be commonly used for analysis of a plurality of target nucleic acids differs in the concentration adjustment reagent tanks 12a and 12b. Concentration adjusting reagents 112a and 112b including the concentration are filled, respectively. Further, the non-common reagent tank 13 is divided into non-common reagent tanks 13a and 13b. Each tank in the cartridge 1 of this embodiment has a common reagent tank 11, concentration adjusting reagent tanks 12a and 12b, and non-common reagent tanks 13a and 13b arranged in this order from one end to the other end. The invention is not limited to this, and each tank can be arranged in any order.

  In the cartridge 1 of the present embodiment, the number of the common reagent tank 11 is one, but the present invention is not limited to this, and may be plural. As a specific example, the number of the common reagent tanks 11 is 1-2. is there. When a plurality of common reagent tanks 11 are included, each common reagent tank may be filled with a common reagent having the same component, or may be filled with a common reagent containing different components. When the common reagent includes the same component, the same component may have the same concentration or a different concentration in each common reagent. The volume of the common reagent tank 11 is not particularly limited, and can be appropriately set according to the volume of the common reagent 111 filled in the common reagent tank 11, for example.

  In the cartridge 1 of the present embodiment, the common reagent 111 filled in the common reagent tank 11 is a liquid reagent, but the present invention is not limited to this and may be a dry reagent. When the common reagent 111 is a liquid reagent, the volume of the common reagent 111 filled in the common reagent tank 11 is not particularly limited, and can be appropriately set according to, for example, the number of target nucleic acids.

  In the cartridge 1 of the present embodiment, there are two concentration adjustment reagent tanks 12, but the present invention is not limited to this, and there may be a plurality, and as a specific example, the number of concentration adjustment reagent tanks 12 is two. ~ 4. The volume of the concentration adjustment reagent tanks 12a and 12b is not particularly limited, and can be appropriately set according to the volume of the concentration adjustment reagents 112a and 112b filled in the concentration adjustment reagent tank 12, for example.

  In the cartridge 1 of the present embodiment, the concentration adjusting reagents 112a and 112b filled in the concentration adjusting reagent tanks 12a and 12b are liquid reagents, but the present invention is not limited to this and may be a dry reagent. When the concentration adjusting reagents 112a and 112b are liquid reagents, the volume of the concentration adjusting reagents 112a and 112b filled in the concentration adjusting reagent tanks 12a and 12b is not particularly limited. For example, depending on the number of the target nucleic acids, Can be set.

  In the cartridge 1 of the present embodiment, the number of non-common reagent tanks 13 is two. However, the present invention is not limited to this, and a plurality of non-common reagent tanks 13 may be used. As a specific example, the number of non-common reagent tanks 13 is two. ~ 4. The volumes of the non-common reagent tanks 13a and 13b are not particularly limited, and can be appropriately set according to the volumes of the non-common reagents 113a and 113b filled in the non-common reagent tank 13, for example.

  In the cartridge 1 of the present embodiment, the non-common reagent tanks 13a and 13b are not filled with the non-common reagent, but the present invention is not limited to this, and the non-common reagent may be filled. When the non-common reagent is filled, the common reagent filled in the non-common reagent tanks 13a and 13b preferably includes different target nucleic acid analysis oligonucleotides. As described above, the non-common reagent containing the analytical oligonucleotide for analyzing the target nucleic acid is prepared in advance and filled in the non-common reagent tanks 13a and 13b, so that the target nucleic acid can be analyzed more easily. it can.

  Next, an analysis method using the cartridge 1 of the present embodiment will be described. First, the non-common reagent tanks 13a and 13b of the cartridge 1 are filled with non-common reagents, respectively. Next, the sample, the common reagent 111 filled in the common reagent tank 11, at least one of the concentration adjusting reagents 112a and 112b filled in the concentration adjusting reagents 12a and 12b, and the non-common reagent tanks 13a and 13b are filled. One non-common reagent is mixed to prepare the first analytical system. Also, the sample, the common reagent 111 filled in the common reagent tank 11, at least one of the concentration adjustment reagents 112a and 112b filled in the concentration adjustment reagents 12a and 12b, and the other non-common reagent are mixed, A second analytical system is prepared.

  Then, the target nucleic acid is analyzed by analyzing the first analysis system and the second analysis system. The method for analyzing the target nucleic acid is not particularly limited, and can be performed by, for example, a known nucleic acid analysis method according to the target nucleic acid. When the target nucleic acid is DNA and the analysis target of the target nucleic acid is SNP, for example, the first analysis system and the second analysis system are analysis oligonucleotides included in the non-common reagent. Using a primer capable of amplifying the target nucleic acid, the target nucleic acid is amplified by a nucleic acid amplification method, and the target nucleic acid and its amplified fragment are melted using a probe capable of detecting the target nucleic acid that is the oligonucleotide for analysis. Analyze by performing a curve analysis.

  The cartridge 1 of this embodiment may further include a sample tank that can be filled with a sample. The numbers of the sample and the sample tank are not particularly limited and are, for example, 1 to 4, respectively. The volume of the sample tank is not particularly limited and can be appropriately determined according to the volume of the sample. The sample is not particularly limited, and examples thereof include biological samples. Examples of the biological sample include blood (for example, blood cells such as whole blood and leukocytes, plasma, serum, etc.), tissue (for example, large intestine, lung, etc.), oral cells (for example, oral mucosa, etc.), somatic cells (for example, , Nails and hair, etc.), germ cells, sputum, amniotic fluid, peritoneal fluid, urine, gastric juice and the like. In addition, examples of the biological sample include biological samples such as gastric lavage fluid, paraffin-embedded tissue, and cultured cells.

  The cartridge 1 of the present embodiment may further include a sample pretreatment reagent and a pretreatment reagent tank filled with the pretreatment reagent. The numbers of the pretreatment reagent and the pretreatment reagent tank are not particularly limited and are, for example, 1 to 4, respectively. Examples of the pretreatment reagent include a reagent that breaks cell membranes such as cells contained in the sample, a reagent that isolates nucleic acids such as DNA and RNA, a reagent that synthesizes DNA from RNA, and the like. Specific examples of the pretreatment reagent include a surfactant, a reducing agent, a protease inhibitor, a DNase inhibitor, an RNase inhibitor, and a reverse transcriptase such as an RNA-dependent DNA polymerase.

  The cartridge 1 of this embodiment may further include other reagents related to the analysis of the target nucleic acid and other reagent tanks filled with the other reagents. The number of the other reagents and other reagent tanks is not particularly limited, and is, for example, 1 to 4, respectively. The other reagents are not particularly limited, and examples thereof include mineral oil and a diluent for diluting each reagent.

(Embodiment 2)
2A and 2B are schematic views of the cartridge of the second embodiment. FIG. 2A is a perspective view of the cartridge of the second embodiment, and FIG. 2B is a view taken in the II-II direction of FIG. It is sectional drawing. In FIG. 2, the same parts as those in FIG. As shown in FIG. 2A, the cartridge 2 further includes a sample tank 14, a pretreatment reagent tank 15, and another reagent tank 16, and a common reagent tank 11, a concentration adjusting reagent tank 12, The upper openings of the non-common reagent tank 13, the pretreatment reagent tank 15, and the other reagent tanks 16 are sealed, and the sample tank 14 is opened at the top. The non-common reagent tanks 13a and 13b are filled with non-common reagents 113a and 113b, respectively. The pretreatment reagent tank 15 is divided into pretreatment reagent tanks 15a and 15b, and the pretreatment reagent tanks 15a and 15b are filled with pretreatment reagents 115a and 115b, respectively. The other reagent tanks 16 are filled with other reagents 116. Except for this point, the cartridge 2 of the present embodiment has the same configuration as the cartridge 1 of the first embodiment, and the description thereof can be used. According to the cartridge 2 of the present embodiment, for example, a target nucleic acid can be analyzed even for a sample that requires pretreatment. Each tank in the cartridge 2 of the present embodiment has another reagent tank 16, a common reagent tank 11, concentration adjusting reagent tanks 12a and 12b, pretreatment reagent tanks 15a and 15b, non-common reagent tanks from one end to the other end. Although 13a, 13b and the sample tank 14 are arrange | positioned in this order, this invention is not limited to this, Each tank can be arrange | positioned in arbitrary orders.

  In the cartridge 2 of this embodiment, the non-common reagent tanks 13a and 13b are filled with non-common reagents 113a and 113b. However, the present invention is not limited to this, and the non-common reagents 113a and 113b are filled. It does not have to be. Further, the non-common reagents 113a and 113b filled in the non-common reagent tanks 13a and 13b are liquid reagents, but the present invention is not limited to this and may be dry reagents. When the non-common reagent 113a, 113b is a liquid reagent, the volume of the non-common reagent 113a, 113b filled in the non-common reagent tank 13a, 13b is not particularly limited. For example, depending on the number of the target nucleic acids, Can be set.

  The cartridge 2 of the present embodiment includes the seal 17, but the present invention is not limited to this, and the seal 17 may have any configuration and may or may not be present. The seal 17 seals the upper openings of the common reagent tank 11, the concentration adjusting reagent tank 12, the non-common reagent tank 13, the pretreatment reagent tank 15, and the other reagent tanks 16. The seal 17 is not limited to the upper opening of any one of the common reagent tank 11, the concentration adjusting reagent tank 12, the non-common reagent tank 13, the sample tank 14, the pretreatment reagent tank 15, and the other reagent tanks 16. It may be sealed, a plurality of upper openings may be sealed, or all the upper openings may be sealed. When the seal 17 seals a plurality of upper openings, the seal 17 may seal, for example, the common reagent tank 11 and the concentration adjustment reagent tank 12, or the common reagent tank 11, the concentration adjustment reagent tank 12, and the front. The processing reagent tank 15 and other reagent tanks 16 may be sealed. The seal 17 is not particularly limited, and a known seal member can be used.

  Next, an analysis method using the cartridge 2 of the present embodiment will be described. First, the sample is filled in the sample tank 14 of the cartridge 2. Next, the sample is divided into two, and for each sample, the sample and at least one of the pretreatment reagents 115a and 115b filled in the pretreatment reagent tanks 15a and 15b are mixed, and the sample is mixed. Perform pre-processing. Then, at least one of the pretreated sample, the common reagent 111 filled in the common reagent tank 11, the concentration adjusting reagents 112a and 112b filled in the concentration adjusting reagents 12a and 12b, and the non-common reagent tank 13a. , 13b are mixed with one of the non-common reagents 113a and 113b, to prepare a first analysis system. The other pretreated sample, the common reagent 111 filled in the common reagent tank 11, at least one of the concentration adjustment reagents 112a and 112b filled in the concentration adjustment reagents 12a and 12b, and the non-common reagent 113a, A second analytical system is prepared by mixing with the other of 113b. Then, another reagent 116 (for example, mineral oil) filled in the other reagent tank 16 is added to the first analysis system and the second analysis system. Then, the first analysis system and the second analysis system are analyzed in the same manner as in the first embodiment.

<Second target nucleic acid analysis reagent cartridge>
As described above, the second cartridge of the present invention has a plurality of common reagent vessels and a plurality of non-common reagent vessels, and each of the common reagent vessels is commonly used for analyzing a plurality of target nucleic acids. The common reagent containing a plurality of components that can be used is filled, and each of the common reagents includes at least one of the plurality of components at different concentrations, and each of the plurality of non-common reagent vessels includes: A non-common reagent containing an analytical oligonucleotide for analyzing a specific target nucleic acid can be filled. The second cartridge of the present invention includes the plurality of common reagent tanks, and each of the common reagents includes at least one component of the plurality of components at different concentrations, and the other configuration The conditions are not particularly limited. In the second cartridge of the present invention, the common reagent contains the adjustment component in the concentration adjusting reagent of the first cartridge of the present invention. For this reason, the second cartridge of the present invention is, for example, a cartridge that includes a plurality of the common reagents of the first cartridge of the present invention, and each of the plurality of common reagents includes the concentration adjusting reagent. You can also. For example, the description of the first cartridge of the present invention can be used for the second cartridge of the present invention.

  As described above, the second cartridge of the present invention has a plurality of common reagent tanks, and each of the plurality of common reagent tanks is at least one component that can be commonly used for analysis of a plurality of target nucleic acids. Are filled with common reagents containing different concentrations. For this reason, for example, when analyzing two target nucleic acids, the adjustment component is set to an optimum concentration according to the type of analysis oligonucleotide for analyzing each target nucleic acid. By mixing the common reagent set to the concentration and a non-common reagent containing the corresponding analytical oligonucleotide, an analysis system suitable for the analytical oligonucleotide can be easily prepared. When analyzing three or more target nucleic acids, one of the adjustment components is set to the highest component concentration or higher among the optimum concentrations according to the type of analysis oligonucleotide for analyzing each target nucleic acid. The other is set to a concentration lower than the lowest component among the optimum concentrations according to the type of the oligonucleotide for analysis for analyzing each target nucleic acid, for example, a common reagent containing these two adjustment components By mixing the sample and the non-common reagent, an analysis system suitable for the analysis oligonucleotide can be easily prepared. Therefore, according to the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Furthermore, according to the second cartridge of the present invention, since the adjustment component can be an analysis system suitable for the analysis oligonucleotide, the target nucleic acid can be analyzed with better analysis accuracy, for example.

  In the second cartridge of the present invention, the common reagent includes a plurality of components that can be commonly used for analysis of a plurality of target nucleic acids, and includes at least one of the plurality of components at different concentrations. The adjustment component can also be referred to as a component whose concentration is adjusted depending on the type of the non-common reagent analysis oligonucleotide used in combination with the common reagent in the analysis of the target nucleic acid, for example. For the adjustment component, for example, the above description can be used. The number of the common reagents may be plural, for example, 2 to 5. Each of the plurality of common reagents may include at least one of the plurality of components at different concentrations, and the concentrations of the other components may be the same or different. When the number of the common reagents is 3 or more, the concentration of the adjustment component in any two common reagents may be the same. The common reagent may be, for example, a liquid reagent or a solid reagent (dry reagent). In the former case, the common reagent is, for example, a reagent in which the common reagent is dissolved, suspended, or mixed in a solvent. When the common reagent is a solid reagent, the concentration of the adjustment component is, for example, the concentration when the cartridge is dissolved, suspended, or mixed in a predetermined amount of solvent.

  Hereinafter, the second cartridge of the present invention and the analysis method using the same will be described in detail with reference to the drawings. However, the present invention is not limited or limited to the following examples. In the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown, unlike the actual case.

(Embodiment 3)
FIG. 3 is a schematic view of the cartridge of the third embodiment, (A) is a perspective view of the cartridge of the third embodiment, and (B) is viewed in the III-III direction of (A). It is sectional drawing. In FIG. 3, the same parts as those in FIG. As shown in FIG. 3A, the cartridge 3 of this embodiment includes a common reagent tank 11 ′ and a non-common reagent tank 13. Further, as shown in FIG. 3B, the common reagent tank 11 ′ is divided into common reagent tanks 11′a and 11′b, and the common reagent tanks 11′a and 11′b include a plurality of the plurality of common reagent tanks 11′a and 11′b. Common reagents 111′a and 111′b each containing a plurality of components that can be commonly used for analysis of the target nucleic acid and at least one of the plurality of components having different concentrations are packed. . Further, the non-common reagent tank 13 is divided into non-common reagent tanks 13a and 13b. Each tank in the cartridge 3 of the present embodiment has common reagent tanks 11′a, 11′b and non-common reagent tanks 13a, 13b arranged in this order from one end to the other end. Without being limited to this, each tank can be arranged in any order.

  In the cartridge 3 of the present embodiment, the number of common reagent tanks 11 ′ is two. However, the present invention is not limited to this, and a plurality of common reagent tanks 11 ′ may be used. As a specific example, the number of common reagent tanks 11 ′ is two. ~ 5. The volumes of the common reagent tanks 11'a and 11'b are not particularly limited, and can be appropriately set according to the volumes of the common reagents 111'a and 111'b filled in the common reagent tank 11 ', for example. The cartridge 3 of the present embodiment may further include any one or more of the above-described sample tank, pretreatment reagent and pretreatment reagent tank, and other reagents and other reagent tanks. The above description can be incorporated.

  Next, an analysis method using the cartridge 3 of the present embodiment will be described. First, the non-common reagent tanks 13a and 13b of the cartridge 3 are filled with the non-common reagent, respectively. Next, a sample, at least one of the common reagents 111′a and 111′b filled in the common reagent tanks 11′a and 11′b, and one non-common reagent filled in the non-common reagent tank 13 Mix to prepare the first analytical system. Further, the sample is mixed with at least one of the common reagents 111′a and 111′b filled in the common reagent tanks 11′a and 11′b and the other non-common reagent, and a second analysis system is prepared. Prepare. Then, the first analysis system and the second analysis system are analyzed in the same manner as in the first embodiment.

<First target nucleic acid analyzer>
As described above, the first analyzer according to the present invention includes a cartridge housing portion in which the first cartridge according to the present invention is housed, a sample, a common reagent filled in the common reagent tank, and the concentration adjustment. A preparation unit for preparing the target nucleic acid analysis system by mixing at least one concentration adjusting reagent filled in a reagent tank and at least one non-common reagent filled in the non-common reagent tank; And an analysis unit for analyzing the target nucleic acid by analyzing the analysis system. The first analyzer of the present invention is characterized in that it includes a cartridge housing portion in which the first cartridge of the present invention is housed, and other configurations and conditions are not particularly limited. For example, the description of the first cartridge of the present invention can be used for the first analyzer of the present invention. According to the first analyzer of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid.

  Hereinafter, the first analyzer of the present invention and the analysis method using the same will be described in detail with reference to the drawings. However, the present invention is not limited or limited to the following examples. In the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown, unlike the actual case.

(Embodiment 4)
FIG. 4 is a block diagram showing the configuration of the analyzer according to this embodiment. As shown in FIG. 4, the analysis device 4 of this embodiment includes a central processing unit (CPU) 41, a sampling pump unit 42 that is a preparation unit, a nozzle drive unit 43, a measurement unit unit 44 that is an analysis unit, and a storage. A device 45, an input device 46, and an output device 47 are provided as main components. The CPU 41 controls the movement of the sampling pump unit 42, the nozzle drive unit 43, and the measurement unit unit 44. The sampling pump unit 42 drives a pump for sampling the sample and each reagent. The nozzle drive unit 43 moves the nozzle to each tank of the cartridge and moves the nozzle up and down. The measurement unit 44 analyzes the reaction of the analysis system in which the sample and each reagent are mixed. Examples of the reaction analysis method include an optical method and an electrochemical method. In the case of an optical method, the measurement unit 44 includes, for example, a light source and a light receiving unit. In the case of an electrochemical method, for example, an electrode is included. The storage device 45 stores analysis condition information, other information, various programs, and the like, and is configured by appropriately combining flash memory, RAM, ROM, and the like. The input device 46 is for inputting information to the analysis device, and includes a keyboard, a key sheet, a terminal such as a USB, a drive such as a CD and a CD-R, and the like. The output device 47 is for outputting analysis results and other information, and examples thereof include a printer and a display. In the present embodiment, each of the above-described constituent elements may be integrated or may be divided into a plurality of devices. For example, you may combine the analyzer provided with the sampling pump unit 42, the nozzle drive unit 43, and the measurement part unit 44, and the personal computer (PC) containing CPU41, the memory | storage device 45, the input device 46, and the output device 47. FIG.

  Next, an analysis method using the analyzer according to the present embodiment will be described by taking the case of using the cartridge according to the second embodiment as an example. First, the sample is filled in the sample tank 14 of the cartridge 2. When the cartridge 2 is set in the analyzer, the nozzle is automatically moved by the nozzle drive unit 43, the sample in the sample tank 14 is sampled by the sampling pump unit 42, and a plurality of analyzes set in the analyzer 4 is performed. Each sample is introduced into a bath (not shown). Then, the nozzle automatically moves, breaks the seal, samples at least one of the pretreatment reagents 115a and 115b, introduces it into the analysis tank, and performs pretreatment. Subsequently, the nozzle automatically moves, breaks the seal, samples the common reagent 111, introduces it into the analysis tank, and further moves the nozzle, breaks the seal, and the concentration adjusting reagents 112a, 112b. Are sampled and introduced into the analysis tank. Then, the nozzle automatically moves, breaks the seal, samples one of the non-common reagents 113a and 113b, introduces it into the analysis tank, and mixes the first analysis system to prepare the sample. React. Further, the sample is reacted after preparing the second analysis system in the same manner except that the other of the non-common reagents 113a and 113b is sampled. When the reaction of the first analysis system and the second analysis system is PCR, the temperature cycle is controlled by the measurement unit 44. After a certain number of cycles of reaction, the probe in the reaction is detected, for example, by optical techniques. Although the case where the cartridge 2 of the second embodiment is used has been described as an example, the cartridge 1 of the first embodiment may be used.

<Second target nucleic acid analyzer>
The second analyzer of the present invention includes a cartridge housing portion in which the second cartridge of the present invention is housed, a sample, at least one common reagent filled in the common reagent tank, and the non-common reagent. A preparation unit that prepares an analysis system for the target nucleic acid by mixing at least one non-common reagent filled in the tank, and an analysis unit that analyzes the target nucleic acid by analyzing the analysis system It is characterized by including. The second analyzer of the present invention is characterized in that it includes a cartridge housing portion in which the second cartridge of the present invention is housed, and other configurations and conditions are not particularly limited. For example, the description of the first and second cartridges of the present invention and the first target nucleic acid analyzer of the present invention can be used for the second analyzer of the present invention. According to the second analyzer of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid.

  The second analyzer of the present invention uses the second cartridge of the present invention instead of the first cartridge of the present invention, and the preparation section is filled with a sample and the common reagent tank at least. The first target nucleic acid analyzer except that the target nucleic acid analysis system is prepared by mixing one common reagent and at least one non-common reagent filled in the non-common reagent tank. And the description can be used.

  The second analyzer of the present invention prepares the analysis system by, for example, mixing the sample, at least two common reagents, and at least one non-common reagent in the preparation unit.

<Analyzing method of first target nucleic acid>
As described above, the first analysis method of the present invention uses the first cartridge of the present invention and fills the sample, the common reagent filled in the common reagent tank, and the concentration adjusting reagent tank. A preparation step of preparing an analysis system of the target nucleic acid by mixing at least one concentration adjusting reagent and at least one non-common reagent filled in the non-common reagent tank; and analyzing the analysis system And an analysis step of analyzing the target nucleic acid. The first analysis method of the present invention is characterized by using the first cartridge of the present invention, and other processes and conditions are not particularly limited. For example, the description of the first cartridge of the present invention and the first analysis apparatus of the present invention can be used for the first analysis method of the present invention. According to the first analysis method of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid.

<Second target nucleic acid analysis method>
The second analysis method of the present invention uses the second cartridge of the present invention and uses a sample, at least one common reagent filled in the common reagent tank, and at least filled in the non-common reagent tank. The method includes a preparation step of preparing an analysis system for the target nucleic acid by mixing one non-common reagent, and an analysis step for analyzing the target nucleic acid by analyzing the analysis system. The second analysis method of the present invention is characterized by using the second cartridge of the present invention, and other processes and conditions are not particularly limited. The second analysis method of the present invention includes, for example, the description of the first and second cartridges of the present invention, the first and second analysis devices of the present invention, and the first analysis method of the present invention. Can be used. According to the second analysis method of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid.

  In the second analysis method of the present invention, for example, in the preparation step, the analysis system is prepared by mixing the sample, at least two common reagents, and at least one non-common reagent.

  Next, examples of the present invention will be described. In addition, this invention is not restrict | limited at all by the following Example.

[Example 1]
In the analysis using different oligonucleotides for analysis of different target nucleic acids, it was confirmed that the magnesium ion and potassium ion concentrations suitable for each analysis oligonucleotide were different.

  As analysis systems for the mutation of codon 117 of the NRAS gene (hereinafter referred to as “NRAS”) and analysis of the mutation of codon 146 of NRAS, the analysis systems having the compositions shown in Tables 1 and 2 below were prepared, respectively. As shown in Table 1 below, the analysis system for NRAS codon 117 includes, as the analysis oligonucleotide, forward primer 1 and reverse primer 1 capable of amplifying a polynucleotide containing a mutation of codon 117 of NRAS, and codon 117 of NRAS. This is an analysis system comprising a probe 1 capable of detecting the mutation 1 and a probe 2 capable of detecting a specific mutation of codon 117 of NRAS. In addition, as shown in Table 2 below, the NRAS codon 146 analysis system uses, as the analysis oligonucleotide, a forward primer 2 and a reverse primer 2 that can amplify a polynucleotide containing a mutation in the codon 146 of NRAS, and an NRAS codon. The analysis system includes a probe 3 capable of detecting a mutation of codon 146. In each analytical system, the concentration of magnesium ions and potassium ions of the adjustment component is adjusted to an optimum concentration for each analytical oligonucleotide.

  Each analysis system was subjected to PCR and melting curve analysis using a gene analyzer (i-densy ™). As a result, in any analysis system, the target mutation could be analyzed with excellent analysis accuracy. From the above, it was found that the magnesium ion and potassium ion concentrations suitable for each oligonucleotide for analysis were different.

[Example 2]
Using two kinds of concentration adjusting reagents each containing different concentrations of magnesium ions, the NRAS gene mutation that is a target nucleic acid can be reacted at a magnesium ion concentration suitable for the oligonucleotide for analyzing the target nucleic acid, and It was confirmed that the target nucleic acid can be analyzed with better analytical accuracy.

(1) Reagents Two types of concentration adjusting reagents 1 and 2 containing a common reagent including DNA polymerase, BSA and the like and different concentrations of magnesium ions were prepared. In addition, non-common reagents 1 to 4 containing dNTP were prepared as non-common reagents. Non-common reagent 1 is a reagent including a primer capable of amplifying a polynucleotide containing mutations of codons 12 and 13 of NRAS, and a probe capable of detecting the mutation of codons 12 and 13 of NRAS. A reagent comprising a primer capable of amplifying a polynucleotide comprising mutations of codons 59 and 61 of NRAS, and a probe capable of detecting the mutations of codons 59 and 61 of NRAS, and non-common reagent 3 comprises a mutation of codon 117 of NRAS A reagent capable of amplifying a polynucleotide containing NRAS and a probe capable of detecting a mutation in codon 117 of NRAS, and non-common reagent 4 is a primer capable of amplifying a polynucleotide containing a mutation in codon 146 of NRAS, As well as pros capable of detecting mutations in codon 146 of NRAS It is a reagent that contains the drive. The concentration adjusting reagent 1 is a reagent in which the concentration of the magnesium ion that is the adjusting component is adjusted according to the primer that is the analytical oligonucleotide of the non-common reagents 1 and 2. The concentration adjusting reagent 2 is a reagent in which the concentration of magnesium ions as the adjusting component is adjusted in accordance with the primer that is the oligonucleotide for analysis of the non-common reagents 3 and 4.

(2) Sample Oligonucleotides containing each mutation were used as samples.

(3) PCR and melting curve analysis Then, each reagent was combined in a predetermined amount to prepare an analytical system, and PCR and melting curve analysis were performed using the gene analyzer. As a result, the target mutation could be analyzed in any combination of reagents. In addition, the non-common reagents 1 and 2 were able to analyze the target mutation with higher accuracy when combined with the concentration adjusting reagent 1 than when combined with the concentration adjusting reagent 2. Furthermore, the non-common reagents 3 and 4 were able to analyze the target mutation with higher accuracy when combined with the concentration adjusting reagent 2 than when combined with the concentration adjusting reagent 1. From these results, it was found that according to the cartridge of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Moreover, according to the cartridge of this invention, it turned out that a target nucleic acid can be analyzed with the outstanding analysis precision.

[Example 3]
Using two types of concentration adjusting reagents each containing different concentrations of magnesium ions, the magnesium ion concentration suitable for the target nucleic acid analysis oligonucleotide for mutation of the target nucleic acid KRAS gene (hereinafter referred to as “KRAS”) It was confirmed that the target nucleic acid can be analyzed with better analytical accuracy.

(1) Reagents Two types of concentration adjusting reagents 1 and 2 containing a common reagent including DNA polymerase, BSA and the like and different concentrations of magnesium ions were prepared. Further, non-common reagents 5 to 8 containing dNTP were prepared as non-common reagents. Non-common reagent 5 is a reagent including a primer capable of amplifying a polynucleotide containing mutations of codons 12 and 13 of KRAS, and a probe capable of detecting mutations of codons 12 and 13 of KRAS. A reagent comprising a primer capable of amplifying a polynucleotide containing mutations of codons 59 and 61 of KRAS, and a probe capable of detecting mutations of codons 59 and 61 of KRAS, and non-common reagent 7 is a mutation of codon 117 of KRAS A reagent capable of amplifying a polynucleotide containing KRAS and a probe capable of detecting a mutation in codon 117 of KRAS, and non-common reagent 8 is a primer capable of amplifying a polynucleotide containing a mutation in codon 146 of KRAS, As well as pros capable of detecting mutations in codon 146 of KRAS It is a reagent that contains the drive. The concentration adjusting reagent 1 is a reagent in which the concentration of magnesium ions as the adjusting component is adjusted according to the primer that is the analytical oligonucleotide of the non-common reagents 5 and 6. The concentration adjusting reagent 2 is a reagent in which the concentration of magnesium ions as the adjusting component is adjusted according to the primer that is the analytical oligonucleotide of the non-common reagents 7 and 8.

(2) Sample Oligonucleotides containing each mutation were used as samples.

(3) PCR and melting curve analysis Then, each reagent was combined in a predetermined amount to prepare an analytical system, and PCR and melting curve analysis were performed using the gene analyzer. As a result, the target mutation could be analyzed in any combination of reagents. In addition, the non-common reagents 5 and 6 were able to analyze the target mutation with higher accuracy when combined with the concentration adjusting reagent 1 than when combined with the concentration adjusting reagent 2. Further, the non-common reagents 7 and 8 were able to analyze the target mutation with higher accuracy when combined with the concentration adjusting reagent 2 than when combined with the concentration adjusting reagent 1. From these results, it was found that according to the cartridge of the present invention, in the analysis of a plurality of target nucleic acids, the reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Moreover, according to the cartridge of this invention, it turned out that a target nucleic acid can be analyzed with the outstanding analysis precision.

  As mentioned above, although this invention was demonstrated with reference to embodiment and an Example, this invention is not limited to the said embodiment and Example. Various changes that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.

  According to the present invention, in analysis of a plurality of target nucleic acids, a reaction can be performed at a component concentration suitable for the oligonucleotide for analysis of each target nucleic acid. Therefore, according to the present invention, for example, the target nucleic acid can be analyzed with better analysis accuracy. Therefore, it can be said that the present invention is extremely useful in, for example, the life science field, the clinical field, and the like.

1, 2, 3 Cartridge 11, 11 ', 11'a, 11'b Common reagent tank 111, 111'a, 111'b Common reagent 12, 12a, 12b Concentration adjusting reagent tank 112, 112a, 112b Concentration adjusting reagent 13 , 13a, 13b Non-common reagent tank 113, 113a, 113b Non-common reagent 14 Sample tank 15, 15a, 15b Pretreatment reagent tank 115, 115a, 115b Pretreatment reagent 16 Other reagent tank 116 Other reagent 17 Seal 4 Analyzer 41 CPU
42 Sampling pump unit 43 Nozzle drive unit 44 Measuring unit 45 Storage device 46 Input device 47 Output device

Claims (15)

Having at least one common reagent tank, a plurality of concentration adjusting reagent tanks, and a plurality of non-common reagent tanks;
Each of the common reagent tanks is filled with a common reagent containing a plurality of components that can be used in common for analysis of a plurality of target nucleic acids,
Each of the plurality of concentration adjustment reagent tanks is filled with a concentration adjustment reagent containing at least one component that can be commonly used for analysis of a plurality of target nucleic acids at different concentrations,
Each of the plurality of non-common reagent tanks can be filled with a non-common reagent containing an analysis oligonucleotide for analyzing a specific target nucleic acid. The components contained at different concentrations in the concentration adjusting reagent,
2. The cartridge according to claim 1, wherein in the analysis of the target nucleic acid, the cartridge is a component whose concentration is adjusted according to the type of oligonucleotide for analysis of the non-common reagent used in combination with the common reagent and the concentration adjusting reagent. . The cartridge according to claim 1, wherein the common reagent does not include a component contained in the concentration adjusting reagent at a different concentration. It has a plurality of common reagent tanks and a plurality of non-common reagent tanks,
Each of the common reagent tanks is filled with a common reagent containing a plurality of components that can be used in common for analysis of a plurality of target nucleic acids,
Each of the common reagents includes at least one component of the plurality of components at different concentrations,
Each of the plurality of non-common reagent tanks can be filled with a non-common reagent containing an analysis oligonucleotide for analyzing a specific target nucleic acid. The components contained at different concentrations in the common reagent,
The cartridge according to claim 4, wherein the concentration of the target nucleic acid is adjusted according to the type of oligonucleotide for analysis of the non-common reagent used in combination with the common reagent in the analysis of the target nucleic acid. The component contained in the different concentration is at least one selected from the group consisting of magnesium ion, potassium ion, manganese ion, sodium ion, and ammonium ion, according to any one of claims 1 to 5. cartridge. The cartridge according to any one of claims 1 to 6, wherein the components contained in the different concentrations are magnesium ions. The cartridge according to claim 1, wherein the analytical oligonucleotide is a primer capable of amplifying the target nucleic acid. The cartridge according to claim 8, wherein the analytical oligonucleotide further comprises a probe capable of detecting the target nucleic acid. The cartridge according to claim 1, wherein the non-common reagent tank is filled with the non-common reagent in advance. The cartridge according to any one of claims 1 to 10, wherein each non-common reagent filled in each non-common reagent tank contains a different target nucleic acid analysis oligonucleotide. A cartridge housing portion in which the cartridge according to any one of claims 1 to 3 and 6 to 11 is housed;
Mixing a sample, a common reagent filled in the common reagent tank, at least one concentration adjusting reagent filled in the concentration adjusting reagent tank, and at least one non-common reagent filled in the non-common reagent tank A preparation unit for preparing an analysis system for the target nucleic acid,
An analysis device for a target nucleic acid, comprising: an analysis unit that analyzes the target nucleic acid by analyzing the analysis system. 13. The analyzer according to claim 12, wherein the preparation unit prepares the analysis system by mixing the sample, at least two concentration adjusting reagents, and at least one non-common reagent. Using the cartridge according to any one of claims 1 to 3 and 6 to 11,
Mixing a sample, a common reagent filled in the common reagent tank, at least one concentration adjusting reagent filled in the concentration adjusting reagent tank, and at least one non-common reagent filled in the non-common reagent tank A preparation step of preparing an analysis system for the target nucleic acid,
An analysis step of analyzing the target nucleic acid by analyzing the analysis system. The analysis method according to claim 14, wherein in the preparation step, the analysis system is prepared by mixing the sample, at least two of the concentration adjusting reagents, and at least one of the non-common reagents.

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