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JP2007189975A - Reaction vessel - Google Patents

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JP2007189975A
JP2007189975A JP2006012794A JP2006012794A JP2007189975A JP 2007189975 A JP2007189975 A JP 2007189975A JP 2006012794 A JP2006012794 A JP 2006012794A JP 2006012794 A JP2006012794 A JP 2006012794A JP 2007189975 A JP2007189975 A JP 2007189975A
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reaction
unit
reagent
detection unit
pcr
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JP5080740B2 (en
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Hiroyuki Kuroki
広幸 黒木
Masaaki Chino
正晃 地野
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Toppan Inc
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Toppan Printing Co Ltd
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Abstract

【課題】複数の収容部を備える反応容器において、様々な実験条件に応じて収容部の個数を容易に変更できるようにする。
【解決手段】基板3の上面に開口して試薬を収容可能な複数の収容部5,7,9を一体的に設けて構成され、少なくとも2つの前記収容部5,7,9が、個別に形成されると共に相互に連結可能に構成されることを特徴とする反応容器1を提供する。また、一方の収容部5が、前記試薬を収容して保存する試薬収容部11を構成し、前記一方の収容部5と連結可能な他方の収容部7,9が、前記試薬を用いて生化学反応を行う反応検出ユニット17を構成することを特徴とする反応容器1を提供する。
【選択図】図1
In a reaction vessel having a plurality of storage units, the number of storage units can be easily changed according to various experimental conditions.
SOLUTION: A plurality of storage portions 5, 7 and 9 that are open on the upper surface of a substrate 3 and can store a reagent are integrally provided, and at least two of the storage portions 5, 7, and 9 are individually provided. Provided is a reaction vessel 1 formed and configured to be mutually connectable. In addition, one storage unit 5 constitutes a reagent storage unit 11 that stores and stores the reagent, and the other storage units 7 and 9 that can be connected to the one storage unit 5 are formed using the reagent. A reaction vessel 1 is provided that constitutes a reaction detection unit 17 that performs a chemical reaction.
[Selection] Figure 1

Description

この発明は、反応容器に関する。   The present invention relates to a reaction vessel.

近年、化学反応やDNA反応、タンパク質反応等の生化学反応において用いられる反応容器としては、板状に形成されたチップの表面に開口するウェル(収容部)を多数形成したものがある(例えば、特許文献1参照。)。これら多数のウェルは、例えば、種々の試薬を予め収容しておく試薬収容部や、上記試薬を分注して生化学反応を行う反応部等として使用することができるようになっている。すなわち、同一の反応容器上で反応実験を行うことが可能とされている。
従来、この種の反応容器は、試薬収容部及び反応部を一体成形して構成されている。
特開2003−70456号公報
In recent years, as a reaction vessel used in a biochemical reaction such as a chemical reaction, a DNA reaction, or a protein reaction, there is one in which a large number of wells (housing portions) opened on the surface of a chip formed in a plate shape (for example, (See Patent Document 1). These many wells can be used as, for example, a reagent storage section that stores various reagents in advance, a reaction section that dispenses the reagents and performs biochemical reactions, and the like. That is, it is possible to perform a reaction experiment on the same reaction vessel.
Conventionally, this type of reaction container is configured by integrally forming a reagent storage part and a reaction part.
JP 2003-70456 A

ところで、上述した反応実験では、その実験条件に応じて使用する試薬の数や生化学反応を行う数が異なるため、試薬収容部や反応部に要するウェルの個数を変更する必要がある。
しかしながら、上記従来の反応容器は試薬収容部及び反応部を一体成形して構成されるため、反応実験の実験条件に適したウェルの個数を備える反応容器を個別に用意する必要があり、反応容器の製造コストが増大する虞がある。また、予めウェルの個数を余分に形成した反応容器を使用することも考えられるが、使用されないウェルが無駄となるため、反応実験に要するコストが高くなる虞がある。
By the way, in the above-described reaction experiment, the number of reagents used and the number of biochemical reactions to be performed differ depending on the experimental conditions, and therefore it is necessary to change the number of wells required for the reagent storage unit and the reaction unit.
However, since the conventional reaction container is formed by integrally forming the reagent container and the reaction part, it is necessary to prepare reaction containers having the number of wells suitable for the experimental conditions of the reaction experiment. There is a risk that the manufacturing cost of the product increases. Although it is conceivable to use a reaction container in which the number of wells is formed in advance, the wells that are not used are wasted, which may increase the cost required for the reaction experiment.

この発明は、上述した事情に鑑みてなされたものであって、様々な実験条件に応じて収容部の個数を容易に変更できる反応容器を提供することを目的としている。   The present invention has been made in view of the above-described circumstances, and an object thereof is to provide a reaction vessel in which the number of storage portions can be easily changed according to various experimental conditions.

上記課題を解決するために、この発明は以下の手段を提案している。
請求項1に係る発明は、基板の上面に開口して試薬を収容可能な複数の収容部を一体的に設けて構成され、少なくとも2つの前記収容部が、個別に形成されると共に相互に連結可能に構成されることを特徴とする反応容器を提案している。
In order to solve the above problems, the present invention proposes the following means.
The invention according to claim 1 is configured by integrally providing a plurality of accommodating portions that are open on the upper surface of the substrate and can accommodate the reagent, and at least two of the accommodating portions are individually formed and connected to each other A reaction vessel characterized by being configured is proposed.

また、請求項2に係る発明は、請求項1に記載の反応容器において、相互に連結可能な複数の前記収容部が、相互に異なる樹脂材料により形成されていることを特徴とする反応容器を提案している。   The invention according to claim 2 is the reaction container according to claim 1, wherein the plurality of accommodating portions that can be connected to each other are formed of different resin materials. is suggesting.

さらに、請求項3に係る発明は、請求項1又は請求項2に記載の反応容器において、一方の収容部が、前記試薬を収容して保存する試薬収容部を構成し、前記一方の収容部と連結可能な他方の収容部が、前記試薬を用いて生化学反応を行う反応検出ユニットを構成することを特徴とする反応容器を提案している。   Furthermore, the invention according to claim 3 is the reaction container according to claim 1 or claim 2, wherein one storage portion constitutes a reagent storage portion that stores and stores the reagent, and the one storage portion The other container that can be connected to the base plate is a reaction detection unit that performs a biochemical reaction using the reagent.

また、請求項4に係る発明は、請求項3に記載の反応容器において、前記反応検出ユニットが、前記試薬収容部から前記試薬を供給した状態で加熱されるPCR部と、光学分析可能な検出部とから構成され、前記PCR部及び前記検出部が、個別に形成されると共に相互に連結可能に形成されることを特徴とする反応容器を提案している。   The invention according to claim 4 is the reaction container according to claim 3, wherein the reaction detection unit is heated with the reagent being supplied from the reagent storage unit, and detection capable of optical analysis. The reaction vessel is characterized in that the PCR unit and the detection unit are formed separately and can be connected to each other.

さらに、請求項5に係る発明は、請求項3に記載の反応容器において、前記反応検出ユニットが、光学分析可能な検出部からなることを特徴とする反応容器を提案している。   Further, the invention according to claim 5 proposes a reaction container according to claim 3, wherein the reaction detection unit comprises a detection unit capable of optical analysis.

請求項1に記載の発明によれば、複数の収容部を相互に連結して基板を構成することにより、反応実験の様々な実験条件に応じて収容部の個数を容易に変更することができる。したがって、従来のように、上記実験条件に適した収容部の個数を有する反応容器を個別に用意する必要が無くなるため、反応容器の製造コストを低く抑えることができる。また、従来のように、上記実験条件によって使用されない余分な収容部が存在することを防止できるため、反応実験に要するコストを低く抑えることもできる。   According to the first aspect of the present invention, by configuring a substrate by interconnecting a plurality of accommodating portions, the number of accommodating portions can be easily changed according to various experimental conditions of the reaction experiment. . Therefore, it is not necessary to separately prepare reaction containers having the number of accommodating portions suitable for the experimental conditions as in the prior art, so that the manufacturing cost of the reaction containers can be kept low. In addition, since it is possible to prevent the presence of an extra storage portion that is not used depending on the experimental conditions as in the prior art, it is possible to reduce the cost required for the reaction experiment.

また、請求項2に記載の発明によれば、反応実験の実験条件として、個別の収容部に光透過性、耐熱性、試薬に対する耐薬品性等の様々な要求特性があったとしても、これら個別の収容部をそれぞれの要求特性に好適な樹脂材料により形成することができるため、反応容器を効率よく使用することが可能となる。   Further, according to the invention described in claim 2, even if there are various required characteristics such as light permeability, heat resistance, and chemical resistance to the reagent in the individual container as the experimental conditions of the reaction experiment, Since the individual accommodating portions can be formed of a resin material suitable for each required characteristic, the reaction vessel can be used efficiently.

さらに、請求項3に記載の発明によれば、同一の反応容器上で反応実験を行うことができる。そして、試薬の数に応じて一方の収容部の個数、及び、反応検出ユニットにおける生化学反応に要する他方の収容部の個数を容易に変更することができるため、効率よく反応実験を行うことが可能となる。   Furthermore, according to the invention described in claim 3, it is possible to perform a reaction experiment on the same reaction vessel. And since the number of one accommodating part and the number of the other accommodating part required for the biochemical reaction in the reaction detection unit can be easily changed according to the number of reagents, the reaction experiment can be performed efficiently. It becomes possible.

また、請求項4及び請求項5に記載の発明によれば、PCR部や検出部を構成する収容部の個数を容易に変更することができるため、さらに効率よく反応実験を行うことが可能となる。   Moreover, according to the invention of Claim 4 and Claim 5, since the number of the accommodating parts which comprise a PCR part and a detection part can be changed easily, it becomes possible to perform reaction experiment more efficiently. Become.

以下、図1及び図2を参照し、本発明の一実施形態に係る反応容器について説明する。
この実施の形態に係る反応容器1は、化学反応やDNA反応、タンパク質反応等の生化学反応を同一のチップ上にて行うμ―Total Analysis System技術やLab−on−Chip技術で利用されるものであり、例えば図1に示すように、縦横寸法が数ミリ角に設定された略長方形で板状の基板3を備えている。この基板3には、その上面に開口して断面略半円状に形成された複数のウェル(収容部)5,7,9が配置されている。
これらウェル5,7,9の配置は、6個の部分、1個の部分、20個の部分に分かれており、夫々用途に応じて試薬収容部11、PCR(ポリメラーゼ連鎖反応)部13、検出部15を構成している。なお、試薬収容部11、PCR部13、検出部15を構成するウェル5,7,9の個数は適宜設定しても良い。
Hereinafter, with reference to FIG.1 and FIG.2, the reaction container which concerns on one Embodiment of this invention is demonstrated.
The reaction container 1 according to this embodiment is used in μ-Total Analysis System technology or Lab-on-Chip technology that performs biochemical reactions such as chemical reactions, DNA reactions, and protein reactions on the same chip. For example, as shown in FIG. 1, a substantially rectangular and plate-like substrate 3 having vertical and horizontal dimensions set to several millimeters square is provided. The substrate 3 is provided with a plurality of wells (accommodating portions) 5, 7, 9 that are open on the upper surface and have a substantially semicircular cross section.
The arrangement of the wells 5, 7, and 9 is divided into six parts, one part, and 20 parts, and a reagent storage unit 11, a PCR (polymerase chain reaction) unit 13, and a detection according to each use. Part 15 is configured. The number of wells 5, 7, and 9 constituting the reagent storage unit 11, the PCR unit 13, and the detection unit 15 may be set as appropriate.

試薬収容部11は、PCR部13で用いる検体試薬やその他の試薬、その後の検出反応に用いる試薬、バッファー、希釈液等の種々の液体(試薬)を注入して保存する収容部となるものであり、この試薬収容部11のウェル5の大きさは、収容される液体の量に応じて適宜設定されている。試薬収容部11のウェル5には、PCR部13や検出部15のウェル7,9よりも長い時間にわたって上記液体を収容するため、液体と反応してウェル5を形成する材料が溶出しないように、また、液体がウェル5に吸着しないように、試薬収容部11のウェル5は溶出性や吸着性が低い材料、すなわち、耐薬品性に優れた材料形成される必要がある。なお、試薬収容部11のウェル5内には、上記液体の他に検体DNAを収容しておいても良い。   The reagent storage unit 11 serves as a storage unit for injecting and storing various liquids (reagents) such as sample reagents and other reagents used in the PCR unit 13, reagents used in subsequent detection reactions, buffers, and diluents. In addition, the size of the well 5 of the reagent storage unit 11 is appropriately set according to the amount of liquid stored. In the well 5 of the reagent storage unit 11, the liquid is stored for a longer time than the wells 7 and 9 of the PCR unit 13 and the detection unit 15, so that the material that forms the well 5 by reacting with the liquid does not elute. In addition, the well 5 of the reagent container 11 needs to be formed of a material with low elution and adsorption properties, that is, a material with excellent chemical resistance so that the liquid does not adsorb to the well 5. In addition to the liquid, sample DNA may be stored in the well 5 of the reagent storage unit 11.

PCR部13は、試薬収容部11と検出部15との間に配置されており、例えば、血液などから抽出したDNAを増幅させるポリメラーゼ連鎖反応等の生化学反応を行うためのものである。なお、この生化学反応ではウェル7を加熱する必要があるため、PCR部13のウェル7は耐熱性に優れている必要がある。
検出部15は、PCR部13で調整した検体DNAをプローブDNAやその他の試薬と反応させることによりDNAの配列を光学分析するところであり、分析するDNA等の数に応じた複数のウェル9を備えている。なお、上記光学分析は、検体DNAあるいは核酸プローブに付けた標識物質(例えば、蛍光物質)の有無を、ウェル9の底面側(基板3の下面側)から検出する発光検出により行われる。したがって、検出部15のウェル9は光透過性に優れている必要となる。これらPCR部13及び検出部15により、上述した各種生化学反応を行う反応検出ユニット17が構成される。
The PCR unit 13 is disposed between the reagent storage unit 11 and the detection unit 15 and performs, for example, a biochemical reaction such as a polymerase chain reaction that amplifies DNA extracted from blood or the like. In this biochemical reaction, since the well 7 needs to be heated, the well 7 of the PCR unit 13 needs to have excellent heat resistance.
The detection unit 15 optically analyzes the DNA sequence by reacting the sample DNA prepared by the PCR unit 13 with probe DNA or other reagents, and includes a plurality of wells 9 corresponding to the number of DNAs to be analyzed. ing. The optical analysis is performed by luminescence detection in which the presence or absence of a labeling substance (for example, a fluorescent substance) attached to the sample DNA or the nucleic acid probe is detected from the bottom surface side of the well 9 (the lower surface side of the substrate 3). Therefore, the well 9 of the detection unit 15 needs to be excellent in light transmittance. The PCR unit 13 and the detection unit 15 constitute a reaction detection unit 17 that performs the various biochemical reactions described above.

これら試薬収容部11、PCR部13及び検出部15の各部は、図2に示すように、それぞれ個別の部材により構成されている。すなわち、基板3が、試薬収容部11のウェル5を含む収容部板材21と、PCR部のウェル7を含むPCR部板材23と、検出部15のウェル9を含む検出部板材25とから構成されている。収容部板材21及びPCR部板材23は、平面視略矩形状に形成されており、検出部板材25に形成された略矩形状の2つの貫通孔25a,25bに基板3の厚さ方向から各々嵌め合わせて、検出部板材25に連結することができるようになっている。   Each part of the reagent storage unit 11, the PCR unit 13, and the detection unit 15 is composed of individual members as shown in FIG. That is, the substrate 3 is composed of a container plate 21 including the well 5 of the reagent container 11, a PCR plate 23 including the well 7 of the PCR unit, and a detector plate 25 including the well 9 of the detector 15. ing. The housing part plate member 21 and the PCR part plate member 23 are formed in a substantially rectangular shape in plan view, and are inserted into two substantially rectangular through holes 25a and 25b formed in the detection unit plate member 25 from the thickness direction of the substrate 3, respectively. It can be fitted and connected to the detection member plate 25.

収容部板材21は、PP(ポリプロピレン)等の耐薬品性に優れた樹脂材料により形成されている。また、PCR部板材23は、耐熱性に優れたPC(ポリカーボネート)、PET(ポリエチレン−テレフタレート)や各種エンジニアリングプラスチック等の樹脂材料により形成されている。さらに、検出部板材25は、PC(ポリカーボネート)、アクリル、シクロオレフィン系ポリマー等の光透過性に優れた樹脂材料により形成されている。すなわち、これら収容部板材21、PCR部板材23及び検出部板材25は、相互に異なる樹脂材料により形成することができるようになっている。   The accommodating part board | plate material 21 is formed with the resin material excellent in chemical resistance, such as PP (polypropylene). Moreover, the PCR part board | plate material 23 is formed with resin materials, such as PC (polycarbonate) excellent in heat resistance, PET (polyethylene terephthalate), and various engineering plastics. Furthermore, the detection part board | plate material 25 is formed with resin materials excellent in light transmittance, such as PC (polycarbonate), an acryl, and a cycloolefin type polymer. That is, the accommodating part plate member 21, the PCR part plate member 23, and the detection part plate member 25 can be formed of different resin materials.

以上のように構成された反応容器1を用いて生化学反応実験を行う方法の一例について、以下に説明する。
はじめに、例えばポリメラーゼ連鎖反応等の各種の反応処理に用いる検体試薬および他の試薬と、検出工程で用いる各種の試薬と、希釈液またはバッファー液等とを、反応容器1の試薬収容部11に収容する試薬収容工程を行う。
次いで、反応容器1のPCR部13のウェル7に反応溶液を供給する反応液供給工程を行う。なお、上記反応溶液は、例えばポリメラーゼ連鎖反応に使用するものであり、血液等から抽出したDNAまたは予め生成された鋳型DNAと、ポリメラーゼ酵素と、各塩基の材料であるdNTP(デオキシヌクレオチド3リン酸)と、pHおよび濃度調整のための希釈液またはバッファー液とからなる。
An example of a method for conducting a biochemical reaction experiment using the reaction vessel 1 configured as described above will be described below.
First, for example, sample reagents and other reagents used in various reaction processes such as polymerase chain reaction, various reagents used in the detection process, and diluents or buffer solutions are accommodated in the reagent container 11 of the reaction vessel 1. A reagent containing step is performed.
Next, a reaction liquid supply step for supplying a reaction solution to the well 7 of the PCR unit 13 of the reaction container 1 is performed. The above reaction solution is used for, for example, the polymerase chain reaction. DNA extracted from blood or the like or template DNA generated in advance, polymerase enzyme, and dNTP (deoxynucleotide triphosphate) which is a material of each base ) And a diluent or buffer solution for adjusting pH and concentration.

その後、PCR部13を加熱するための温度制御装置(不図示)を、反応容器1のPCR部13を基板3の上面及び下面から挟み込むようにして配置する。なお、上記温度制御装置は例えばペルチェ素子等により構成される。そして、この状態においてポリメラーゼ連鎖反応を生じさせる反応生成工程を行う。
すなわち、この反応生成工程においては、はじめに、温度制御装置によりPCR部13の温度状態を、所定時間(例えば、5〜25秒等)に亘って、所定温度(例えば、90〜100℃程度)となるように制御し、反応溶液のDNAを熱変性させる変性工程を行う。
次いで、温度制御装置によりPCR部13の温度状態を、所定時間(例えば、15〜60秒等)に亘って、所定温度(例えば、50〜60℃程度)となるように制御し、各種のプライマー(つまり、DNAの断片)を所望の遺伝子配列と結合(アニーリング)させるアニーリング工程を行う。
Thereafter, a temperature control device (not shown) for heating the PCR unit 13 is arranged so as to sandwich the PCR unit 13 of the reaction vessel 1 from the upper surface and the lower surface of the substrate 3. In addition, the said temperature control apparatus is comprised by the Peltier element etc., for example. And in this state, the reaction production | generation process which produces a polymerase chain reaction is performed.
That is, in this reaction generation step, first, the temperature state of the PCR unit 13 is set to a predetermined temperature (for example, about 90 to 100 ° C.) over a predetermined time (for example, 5 to 25 seconds) by the temperature controller. The denaturation process which heat-denatures the DNA of a reaction solution is performed.
Next, the temperature control device controls the temperature state of the PCR unit 13 to be a predetermined temperature (for example, about 50 to 60 ° C.) for a predetermined time (for example, 15 to 60 seconds), and various primers In other words, an annealing step is performed in which (ie, a DNA fragment) is combined (annealed) with a desired gene sequence.

その後、温度制御装置によりPCR部13の温度状態を、所定時間(例えば、1〜5分等)に亘って、所定温度(例えば、65〜75℃程度)となるように制御し、DNAポリメラーゼによる相補鎖合成を行う伸長反応工程を行う。
この伸長反応工程の終了後には、上述した変性工程から伸長反応工程までの一連の処理を継続するか否かを判定し、継続する場合には再度変性工程から一連の処理を行う。一方、一連の処理を継続しないと判定された場合には、反応生成工程を終了する。
Thereafter, the temperature state of the PCR unit 13 is controlled by the temperature control device so as to reach a predetermined temperature (for example, about 65 to 75 ° C.) for a predetermined time (for example, 1 to 5 minutes, etc.). An extension reaction step for synthesizing complementary strands is performed.
After the end of the extension reaction step, it is determined whether or not to continue the series of processes from the above-described denaturation step to the extension reaction step. If so, the series of processing is performed again from the denaturation step. On the other hand, when it is determined not to continue the series of processes, the reaction generation step is terminated.

反応生成工程の終了後には、温度制御装置を反応容器1のPCR部13から離間させ、その後に、生成された反応生成物をPCR部13から回収する回収工程を行う。
最後に、反応生成工程でのポリメラーゼ連鎖反応によって調整された検体と、検出用の各種の試薬(例えば、核酸プローブ等)とを、反応容器1の検出部15においてハイブリダイゼーション等により反応させ、予め検体あるいは核酸プローブに付けた標識物質(例えば、蛍光物質)の有無を、反応容器1の検出部15の下面側から検出する発光検出工程を行い、一連の処理を終了する。
以上説明したように、この反応容器1は、試薬収容部11とPCR部13と検出部15とから構成されるため、PCRによる検体の調製からDNAの分析まで同一チップ上で連続して行うことができる。
After completion of the reaction generation step, the temperature control device is separated from the PCR unit 13 of the reaction vessel 1, and thereafter, a recovery step of recovering the generated reaction product from the PCR unit 13 is performed.
Finally, the sample prepared by the polymerase chain reaction in the reaction generation step and various detection reagents (for example, nucleic acid probes) are reacted in the detection unit 15 of the reaction container 1 by hybridization or the like, A luminescence detection step is performed to detect the presence or absence of a labeling substance (for example, a fluorescent substance) attached to the specimen or nucleic acid probe from the lower surface side of the detection unit 15 of the reaction container 1, and the series of processes is completed.
As described above, since the reaction container 1 is composed of the reagent storage unit 11, the PCR unit 13, and the detection unit 15, the steps from the preparation of the sample by PCR to the analysis of DNA are continuously performed on the same chip. Can do.

なお、反応容器1を利用する生化学反応実験は、上述したものの他に、例えば抗原抗体反応及びDNA反応の検出などに用いることもできる。
抗原抗体反応による抗原検出の場合、例えば、予めPCR部13内に抗原を含む試料を入れておき、後から抗体を含む試薬を添加し、抗原または抗体に標識物質を付けておくことで、反応の有無を検出できる。標識物質としては、蛍光などの発光物質が一般的に用いられる。
In addition, the biochemical reaction experiment using the reaction container 1 can be used for, for example, detection of an antigen-antibody reaction and a DNA reaction in addition to those described above.
In the case of antigen detection by antigen-antibody reaction, for example, a sample containing an antigen is placed in the PCR unit 13 in advance, a reagent containing an antibody is added later, and a labeling substance is attached to the antigen or antibody. The presence or absence of can be detected. As the labeling substance, a luminescent substance such as fluorescence is generally used.

また、DNAの検出の場合、例えば、予め検出部15内に核酸プローブを用意しておく。次に、検体DNAを検出部15のウェル9に供給し、核酸プローブと検体DNAとのハイブリダイゼーション反応により、DNAの検出を行うことができる。その際、検体DNAに標識物質を付けておけば、その標識物質の有無を検出することにより検出が可能となる。
ここで、検体DNAとしては、血液等から抽出したDNAをPCR法、LAMP法などにより調整しておいたものを用いることができる。また、核酸プローブとして配列の異なる核酸を複数用意することで検体DNAがどのような配列であるかを検出することができる。
In the case of detecting DNA, for example, a nucleic acid probe is prepared in the detection unit 15 in advance. Next, the sample DNA can be supplied to the well 9 of the detection unit 15 and the DNA can be detected by a hybridization reaction between the nucleic acid probe and the sample DNA. At this time, if a labeling substance is attached to the sample DNA, detection can be performed by detecting the presence or absence of the labeling substance.
Here, as the sample DNA, DNA extracted from blood or the like and prepared by the PCR method, the LAMP method, or the like can be used. Further, by preparing a plurality of nucleic acids having different sequences as nucleic acid probes, it is possible to detect the sequence of the sample DNA.

さらに、反応容器1は一塩基遺伝子多型(SNP)の解析にも用いることができる。なお、その場合、プローブ核酸やその検出に用いる物質は複数あってもよく、それらの物質のひとつが標識されていればよい。
また、標識物質は、結合したプローブ核酸と検体DNAに特異的に作用するものを、反応後に加えることもできる。このようなものとしては、インターカレーターなどがある。また、ここでいう標識物質とは間接的なものも含む。すなわち、蛍光物質などに結合する物質を標識物質として検体DNAに結合させておき、後から蛍光物質を加えても良い。
Furthermore, the reaction vessel 1 can also be used for analysis of single nucleotide polymorphism (SNP). In that case, there may be a plurality of probe nucleic acids and substances used for the detection, and one of these substances only needs to be labeled.
In addition, as the labeling substance, a substance that specifically acts on the bound probe nucleic acid and the sample DNA can be added after the reaction. Such a thing includes an intercalator. Further, the labeling substance here includes indirect substances. That is, a substance that binds to a fluorescent substance or the like may be bound to the sample DNA as a labeling substance, and the fluorescent substance may be added later.

また、多段階反応を行って上述したSNPまたはDNAを検出してもよい。例えば、インベーダー・アッセイ法(サードウェイブテクノロジーズ,Inc(米国ウィスコンシン州マディソン市)を用いても良い。これによりSNP解析の具現化を図ることが可能となる。
この場合、検出DNAの検出に用いるプローブ核酸などの物質が複数種でもよく、予めPCR部13内に少なくとも1種の物質を入れておき、その後、検出DNAと他の物質を同時または順次注入し、反応をおこなっても良い。
Moreover, you may detect SNP or DNA mentioned above by performing multistep reaction. For example, an invader assay method (Third Wave Technologies, Inc. (Madison, Wisconsin, USA)) may be used, thereby enabling realization of SNP analysis.
In this case, there may be a plurality of types of substances such as probe nucleic acids used for detecting the detection DNA. At least one type of substance is put in the PCR unit 13 in advance, and then the detection DNA and other substances are injected simultaneously or sequentially. , You may react.

上記のように、この反応容器1によれば、収容部板材21、PCR部板材23及び検出部板材25を嵌め合わせにより相互に連結して基板3を構成しているため、同一の反応容器1上で生化学反応実験を行うことができる。そして、収容部板材21、PCR部板材23及び検出部板材25の組み合わせを変更するだけで、生化学反応実験の様々な実験条件に応じて試薬収容部11、PCR部13、検出部15をそれぞれ構成するウェル5,7,9の個数を容易に変更することが可能となる。   As described above, according to this reaction container 1, the container 3, the PCR part plate 23, and the detection part plate 25 are connected to each other by fitting to form the substrate 3. Biochemical reaction experiments can be performed on the above. Then, only by changing the combination of the storage unit plate material 21, the PCR unit plate material 23, and the detection unit plate material 25, the reagent storage unit 11, the PCR unit 13, and the detection unit 15 are respectively changed according to various experimental conditions of the biochemical reaction experiment. It becomes possible to easily change the number of wells 5, 7 and 9 to be configured.

なお、試薬収容部11のウェル5の個数は、例えば生化学反応実験に使用する試薬の数に応じて変更すればよい。また、PCR部13を構成するウェル7の個数は、例えばポリメラーゼ連鎖反応等の生化学反応を行う数に応じて変更すればよい。さらに、検出部15を構成するウェル9の個数は、例えば分析するDNA等の数に応じて変更すればよい。
したがって、この反応容器1によれば、従来のように、上記実験条件に適したウェルの個数を有する反応容器を個別に用意する必要が無くなるため、反応容器1の製造コストを低く抑えることができる。また、従来のように、上記実験条件によって使用されない余分なウェルが存在することも防止できるため、生化学反応実験に要するコストを低く抑えると共に効率よく生化学反応実験を行うことができる。
The number of wells 5 in the reagent storage unit 11 may be changed according to the number of reagents used in the biochemical reaction experiment, for example. The number of wells 7 constituting the PCR unit 13 may be changed according to the number of biochemical reactions such as polymerase chain reaction. Furthermore, the number of wells 9 constituting the detection unit 15 may be changed according to the number of DNAs to be analyzed, for example.
Therefore, according to this reaction container 1, since it is not necessary to separately prepare reaction containers having the number of wells suitable for the experimental conditions as in the prior art, the manufacturing cost of the reaction container 1 can be kept low. . Moreover, since it is possible to prevent the existence of extra wells that are not used depending on the experimental conditions as in the prior art, the cost required for the biochemical reaction experiment can be kept low and the biochemical reaction experiment can be performed efficiently.

また、収容部板材21、PCR部板材23及び検出部板材25は、それぞれ耐薬品性、耐熱性、光透過性等の要求特性に好適な樹脂材料により形成されているため、同一種類の反応容器1を様々な実験条件下における生化学反応実験に利用することも可能となる。すなわち、反応容器1をさらに効率よく使用することが可能となる。なお、ここでの実験条件とは、例えば反応溶液や反応生成物等の種類、温度制御装置による加熱温度、検体の分析方法等のことを示している。   Moreover, since the accommodating part board | plate material 21, the PCR part board | plate material 23, and the detection part board | plate material 25 are each formed of the resin material suitable for required characteristics, such as chemical resistance, heat resistance, and light transmittance, it is the same kind of reaction container. 1 can be used for biochemical reaction experiments under various experimental conditions. That is, the reaction vessel 1 can be used more efficiently. Here, the experimental conditions indicate, for example, the types of reaction solutions and reaction products, the heating temperature by the temperature control device, the analysis method of the specimen, and the like.

なお、上記の実施形態において、収容部板材21及びPCR部板材23は、基板3の厚さ方向から検出部板材25に嵌め合わせるとしたが、これに限ることはない。すなわち、例えば図3,4に示すように、検出部板材35にその側方に開口する切欠部35aを形成しておき、PCR部板材33及び収容部板材31を順次検出部板材35の側部からスライドさせて切欠部35a内に挿入することで、検出部板材35に嵌め合わせるとしても構わない。この構成の場合でも、上記実施形態と同様の効果を奏する。
また、基板3は、収容部板材21,31及びPCR部板材23,33を検出部板材25,35に嵌め合わせて構成されるとしたが、これに限ることはなく、少なくとも収容部板材21,31、PCR部板材23,33及び検出部板材25,35を相互に連結するように構成されていればよい。
In the above-described embodiment, the accommodation unit plate material 21 and the PCR unit plate material 23 are fitted to the detection unit plate material 25 from the thickness direction of the substrate 3, but the present invention is not limited thereto. That is, for example, as shown in FIGS. 3 and 4, a notch 35 a that opens to the side of the detection unit plate 35 is formed in the detection unit plate 35, and the PCR unit plate 33 and the storage unit plate 31 are sequentially arranged on the side of the detection unit plate 35. It is possible to fit the detection part plate material 35 by sliding the sheet from the position and inserting it into the cutout part 35a. Even in the case of this configuration, the same effects as those of the above-described embodiment are obtained.
In addition, the substrate 3 is configured by fitting the accommodation unit plate members 21 and 31 and the PCR unit plate members 23 and 33 to the detection unit plate members 25 and 35, but is not limited thereto. 31, the PCR part board | plate materials 23 and 33 and the detection part board | plate materials 25 and 35 should just be comprised so that it may mutually connect.

さらに、PCR部板材23,33及び検出部板材25,35は、相互に異なる樹脂材料によりそれぞれ形成されることに限らず、PC等のように耐熱性及び光透過性に優れている樹脂材料であれば、同一種類の樹脂材料により形成されるとしてもよいし、同一の樹脂材料により一体的に形成されるとしても構わない。
また、収容部板材21,31、PCR部板材23,33及び検出部板材25,35は、相互に異なる樹脂材料により形成されるとしたが、これら各部材に個別の要求特性(例えば、耐薬品性、耐熱性、光透過性等)が無い場合、若しくは同特性の要求が低い場合には、例えば、同一種類の樹脂材料により形成されるとしても構わない。
Furthermore, the PCR unit plate members 23 and 33 and the detection unit plate members 25 and 35 are not limited to being formed of resin materials different from each other, but are resin materials having excellent heat resistance and light transmission properties such as PC. If it exists, it may be formed of the same type of resin material, or may be integrally formed of the same resin material.
In addition, although the accommodating portion plate materials 21, 31, the PCR portion plate materials 23, 33 and the detection portion plate materials 25, 35 are formed of mutually different resin materials, individual required characteristics (for example, chemical resistance) In the case where there is no property, heat resistance, light transmissivity, or the like, or when the requirement for the same characteristics is low, for example, they may be formed of the same type of resin material.

さらに、反応検出ユニット17はPCR部13を備えるとしたが、DNAを増幅させるポリメラーゼ連鎖反応等の反応が不要である場合には、検出部15のみにより構成されるとしても構わない。この構成の場合でも、これまで述べてきたように、試薬収容部11及び検出部15を、相互に連結して一体的に固定することができるため、試薬収容部11及び検出部15をそれぞれ構成するウェル5,9の個数を容易に変更することができる。
なお、反応容器は、例えば試薬収容部11及びPCR部13のみから構成されるとしてもよく、この場合でも前述と同様の効果を奏することができる。
Furthermore, although the reaction detection unit 17 includes the PCR unit 13, it may be configured by only the detection unit 15 when a reaction such as a polymerase chain reaction for amplifying DNA is not required. Even in this configuration, as described above, since the reagent storage unit 11 and the detection unit 15 can be connected to each other and fixed integrally, the reagent storage unit 11 and the detection unit 15 are respectively configured. The number of wells 5 and 9 to be changed can be easily changed.
Note that the reaction container may be composed of only the reagent storage unit 11 and the PCR unit 13, for example, and in this case, the same effects as described above can be obtained.

以上、本発明の実施形態について図面を参照して詳述したが、具体的な構成はこの実施形態に限られるものではなく、本発明の要旨を逸脱しない範囲の設計変更等も含まれる。   As mentioned above, although embodiment of this invention was explained in full detail with reference to drawings, the concrete structure is not restricted to this embodiment, The design change etc. of the range which does not deviate from the summary of this invention are included.

この発明の一実施形態に係る反応容器を示す概略斜視図である。It is a schematic perspective view which shows the reaction container which concerns on one Embodiment of this invention. 図1の反応容器を構成する収容部板材、PCR部板材及び検出部板材を示す分解斜視図である。It is a disassembled perspective view which shows the accommodating part board | plate material, PCR part board | plate material, and detection part board | plate material which comprise the reaction container of FIG. この発明の他の実施形態に係る反応容器を示す概略斜視図である。It is a schematic perspective view which shows the reaction container which concerns on other embodiment of this invention. 図3の反応容器を構成する収容部板材、PCR部板材及び検出部板材を示す分解斜視図である。It is a disassembled perspective view which shows the accommodating part board | plate material, PCR part board | plate material, and detection part board | plate material which comprise the reaction container of FIG.

符号の説明Explanation of symbols

1 反応容器
3 基板
5,7,9 ウェル(収容部)
11 試薬収容部
13 PCR部
15検出部
17 反応検出ユニット

1 Reaction vessel 3 Substrate 5, 7, 9 well (container)
11 Reagent storage unit 13 PCR unit 15 detection unit 17 reaction detection unit

Claims (5)

基板の上面に開口して試薬を収容可能な複数の収容部を一体的に設けて構成され、
少なくとも2つの前記収容部が、個別に形成されると共に相互に連結可能に構成されることを特徴とする反応容器。
It is configured by integrally providing a plurality of accommodating portions that are open on the upper surface of the substrate and can accommodate the reagent,
The reaction container, wherein at least two of the accommodating portions are formed individually and are connectable to each other.
相互に連結可能な複数の前記収容部が、相互に異なる樹脂材料により形成されていることを特徴とする請求項1に記載の反応容器。   The reaction container according to claim 1, wherein the plurality of accommodating portions that can be connected to each other are formed of different resin materials. 一方の収容部が、前記試薬を収容して保存する試薬収容部を構成し、
前記一方の収容部と連結可能な他方の収容部が、前記試薬を用いて生化学反応を行う反応検出ユニットを構成することを特徴とする請求項1又は請求項2に記載の反応容器。
One storage unit constitutes a reagent storage unit that stores and stores the reagent,
3. The reaction container according to claim 1, wherein the other housing part that can be connected to the one housing part constitutes a reaction detection unit that performs a biochemical reaction using the reagent. 4.
前記反応検出ユニットが、前記試薬収容部から前記試薬を供給した状態で加熱されるPCR部と、光学分析可能な検出部とから構成され、
前記PCR部及び前記検出部が、個別に形成されると共に相互に連結可能に形成されることを特徴とする請求項3に記載の反応容器。
The reaction detection unit is composed of a PCR unit heated with the reagent supplied from the reagent storage unit, and a detection unit capable of optical analysis,
The reaction container according to claim 3, wherein the PCR unit and the detection unit are formed separately and can be connected to each other.
前記反応検出ユニットが、光学分析可能な検出部からなることを特徴とする請求項3に記載の反応容器。

The reaction container according to claim 3, wherein the reaction detection unit includes a detection unit capable of optical analysis.

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