JP2017192310A - Mixing device, and nucleic acid purification method - Google Patents

Mixing device, and nucleic acid purification method Download PDF

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JP2017192310A
JP2017192310A JP2016082988A JP2016082988A JP2017192310A JP 2017192310 A JP2017192310 A JP 2017192310A JP 2016082988 A JP2016082988 A JP 2016082988A JP 2016082988 A JP2016082988 A JP 2016082988A JP 2017192310 A JP2017192310 A JP 2017192310A
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mixing chamber
chamber
mixing
piston
storage chamber
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泰生 隅田
Yasuo Sumida
泰生 隅田
圭佑 合田
Keisuke Aida
圭佑 合田
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HILLTOP
Hilltop Corp
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Hilltop Corp
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Abstract

PROBLEM TO BE SOLVED: To provide a mixing device and a nucleic acid purification method which are capable of easily mixing a plurality of substances.SOLUTION: A nucleic acid purification kit 1 being a mixing device comprises: a base member 10 having a mixing chamber 11; a rotary member 30 rotatably provided for the base member 10; and a piston member 40 slidably provided for the base member 10. A plurality of storage chambers for storing substances are formed on a rotary member 30, the storage chambers are constituted so as to be communicated with the mixing chamber 11 when a rotation angle of the rotary member 30 is at a specific angle, and a communication state between the storage chamber and the mixing chamber 11 switches by changing the rotation angle of the rotary member 30. Further, the piston member 40 is equipped with a piston head 41 that closes the mixing chamber 11, and volume inside the mixing chamber 11 changes by sliding the piston member 40.SELECTED DRAWING: Figure 11

Description

本発明は、例えば検体に含まれるウイルスを検査するために複数の物質を混合する混合器具および当該混合器具を用いた核酸精製方法に関するものである。   The present invention relates to a mixing device for mixing a plurality of substances, for example, for testing a virus contained in a specimen, and a nucleic acid purification method using the mixing device.

唾液等の検体検査の際に、複数の物質を順番に混合することが行われており、得られた混合物を処理することによって、ウイルスの核酸を精製することが知られている。例えば、ウイルスを含む唾液と、糖鎖固定化磁性金属ナノ粒子と、当該金属ナノ粒子よりも平均粒子径が大きい磁性体とを混合し、磁力で吸引して得られた沈殿物と希釈用の水とを混合し、加熱した後にウイルスの核酸を含んだ上清を得る核酸精製方法が存在する（特許文献１参照）。   It is known that a plurality of substances are mixed in order at the time of examination of a sample such as saliva, and the nucleic acid of the virus is purified by processing the obtained mixture. For example, saliva containing virus, sugar chain-immobilized magnetic metal nanoparticles, and a magnetic substance having an average particle size larger than that of the metal nanoparticles are mixed, and the precipitate obtained by suction with magnetic force is diluted with There is a nucleic acid purification method for obtaining a supernatant containing viral nucleic acid after mixing with water and heating (see Patent Document 1).

特開２０１１−４５３５８号公報JP 2011-45358 A

ところで、上記のような複数の物質の混合は、例えばマイクロチューブやピペット等の周知の器具を用いて行うことができるが、混合時に物質が零れないように器具の取り扱いに細心の注意を払う必要があるため、簡単に混合することができなかった。   By the way, mixing of a plurality of substances as described above can be performed using a well-known instrument such as a micro tube or a pipette, but it is necessary to pay close attention to the handling of the instrument so that the substance does not spill during mixing. So it could not be mixed easily.

本発明は、上記事情に鑑みてなされたものであって、複数の物質を簡単に混合することが可能な混合器具および核酸精製方法を提供することを課題とする。   This invention is made | formed in view of the said situation, Comprising: It aims at providing the mixing instrument and nucleic acid purification method which can mix a some substance easily.

上記課題を解決するため、請求項１に記載の混合器具は、複数の物質を混合する混合器具であって、混合室を有したベース部材と、前記ベース部材に対して回転可能に設けられた回転部材と、前記ベース部材に対して摺動可能に設けられたピストン部材とを備え、前記回転部材は、前記物質を貯留するための複数の貯留室を有し、前記貯留室は、前記回転部材の回転角が特定の角度であるとき前記混合室と連通するように構成されており、前記回転部材の回転角を変化させることで、前記貯留室と前記混合室との連通状態が切り替わり、前記ピストン部材は、前記混合室を閉塞するピストンヘッドを有しており、前記ピストン部材を摺動させることで、前記混合室の内部の容積が変化することを特徴とする。   In order to solve the above-mentioned problem, the mixing device according to claim 1 is a mixing device for mixing a plurality of substances, and is provided so as to be rotatable with respect to the base member having a mixing chamber and the base member. A rotating member; and a piston member slidably provided with respect to the base member, wherein the rotating member has a plurality of storage chambers for storing the substance, and the storage chamber is configured to rotate the rotating member. It is configured to communicate with the mixing chamber when the rotation angle of the member is a specific angle, and the communication state between the storage chamber and the mixing chamber is switched by changing the rotation angle of the rotating member, The piston member has a piston head that closes the mixing chamber, and the volume inside the mixing chamber is changed by sliding the piston member.

請求項２に記載の混合器具は、請求項１に記載の混合器具において、前記混合室の内部に磁場を発生させる磁石部材をさらに備え、前記磁石部材は、前記ベース部材に対して摺動可能に設けられていることを特徴とする。   The mixing device according to claim 2 is the mixing device according to claim 1, further comprising a magnet member that generates a magnetic field inside the mixing chamber, wherein the magnet member is slidable with respect to the base member. It is provided in.

請求項３に記載の混合器具は、請求項１または２に記載の混合器具において、前記ピストン部材の摺動を制限する摺動制限部材をさらに備え、前記ピストン部材は、前記ベース部材の内部から外部に延びたピストンロッドを有し、当該ピストンロッドには、当該ピストンロッドが延びる方向において複数の突部が所定間隔をあけて設けられており、前記摺動制限部材は、前記突部に当接することで、前記ピストン部材の摺動を制限することを特徴とする。   The mixing instrument according to claim 3 is the mixing instrument according to claim 1 or 2, further comprising a sliding limiting member that limits sliding of the piston member, wherein the piston member is provided from the inside of the base member. The piston rod has an outwardly extending piston rod, and the piston rod is provided with a plurality of protrusions at a predetermined interval in the direction in which the piston rod extends, and the sliding restriction member contacts the protrusion. By contacting, the sliding of the piston member is limited.

請求項４に記載の混合器具は、請求項１〜３のいずれか一項に記載の混合器具において、前記混合室を閉塞する膜部材と、前記膜部材を穿つことが可能な排出用ノズルとをさらに備え、前記排出用ノズルは、前記膜部材が穿たれることで形成される排出孔に取り付けられることを特徴とする。   A mixing device according to claim 4 is the mixing device according to any one of claims 1 to 3, wherein a membrane member that closes the mixing chamber, and a discharge nozzle that can pierce the membrane member; The discharge nozzle is attached to a discharge hole formed when the membrane member is pierced.

請求項５に記載の核酸精製方法は、前記回転部材が少なくとも２つの前記貯留室を有し、ウイルスを捕捉する糖鎖固定化磁性粒子が前記貯留室としての第１貯留室に貯留され、ウイルスを不活化する殺ウイルス剤が前記貯留室としての第２貯留室に貯留され、前記糖鎖固定化磁性粒子よりも平均粒子径の大きい磁性粒子が前記混合室に貯留されている請求項１〜４のいずれか一項に記載の混合器具を用いた核酸精製方法であって、ウイルスを含んだ検体を前記第１貯留室に入れる第１工程と、前記第１工程の後に、前記第１貯留室と前記混合室とが連通状態となるように前記回転部材を回転させる第２工程と、前記第２工程の後に、前記混合室の内部の容積が拡大するように前記ピストン部材を摺動させることで、前記検体に含まれる前記ウイルスを捕捉した前記糖鎖固定化磁性粒子を、前記第１貯留室から前記混合室に吸入する第３工程と、前記第３工程の後に、前記混合室の内部に磁場を発生させた状態で、前記混合室の内部の容積が縮小するように前記ピストン部材を摺動させることで、前記混合室内の上清を前記第１貯留室に吐出する第４工程と、前記第４工程の後に、前記第２貯留室と前記混合室とが連通状態となるように前記回転部材を回転させる第５工程と、前記第５工程の後に、前記混合室の内部の容積が拡大するように前記ピストン部材を摺動させることで、前記殺ウイルス剤を、前記第２貯留室から前記混合室に吸入する第６工程と、前記第６工程の後に、前記混合室の内部に磁場を発生させた状態で、前記混合室の内部の容積が縮小するように前記ピストン部材を摺動させることで、前記ウイルスの核酸を前記混合室から排出する第７工程とを含むことを特徴とする。   The nucleic acid purification method according to claim 5, wherein the rotating member has at least two storage chambers, and sugar chain-immobilized magnetic particles for capturing a virus are stored in a first storage chamber as the storage chamber. The virus-killing agent which inactivates is stored in the second storage chamber as the storage chamber, and magnetic particles having an average particle size larger than the sugar chain-immobilized magnetic particles are stored in the mixing chamber. 5. A method for purifying nucleic acid using the mixing instrument according to claim 4, wherein a first step of putting a specimen containing a virus into the first storage chamber, and the first storage after the first step. A second step of rotating the rotating member so that the chamber and the mixing chamber are in communication with each other; and after the second step, the piston member is slid so as to increase the internal volume of the mixing chamber Thus, the cough contained in the specimen is In a state where a magnetic field is generated inside the mixing chamber after the third step of sucking the sugar chain-immobilized magnetic particles that have captured the ruth into the mixing chamber from the first storage chamber, and after the third step. The fourth step of discharging the supernatant in the mixing chamber into the first storage chamber by sliding the piston member so that the internal volume of the mixing chamber is reduced, and after the fourth step, A fifth step of rotating the rotating member so that the second storage chamber and the mixing chamber are in communication with each other; and the piston member so that an internal volume of the mixing chamber is expanded after the fifth step. In a state where a magnetic field is generated inside the mixing chamber after the sixth step and the sixth step of sucking the virusicide from the second storage chamber into the mixing chamber. The piston so that the internal volume of the mixing chamber is reduced. By sliding the timber, characterized in that it comprises a seventh step of discharging the nucleic acid of the virus from the mixing chamber.

本発明によれば、複数の物質を簡単に混合することが可能な混合器具および核酸精製方法を提供することができる。   ADVANTAGE OF THE INVENTION According to this invention, the mixing instrument and nucleic acid purification method which can mix a some substance easily can be provided.

本発明の一実施形態に係る混合器具としての核酸精製キットの斜視図である。It is a perspective view of the nucleic acid purification kit as a mixing instrument concerning one embodiment of the present invention. 同実施形態に係る核酸精製キットを構成する器具本体の分解斜視図である。It is a disassembled perspective view of the instrument main body which comprises the nucleic acid purification kit which concerns on the same embodiment. 同実施形態に係る核酸精製キットが備えるベース部材の図であり、（Ａ）は前面図、（Ｂ）は後面図、（Ｃ）は上面図、（Ｄ）は下面図、（Ｅ）は左側面図、（Ｆ）は断面図である。It is a figure of the base member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) is a front view, (B) is a rear view, (C) is a top view, (D) is a bottom view, (E) is a left side. A plane view and (F) are sectional views. 同実施形態に係る核酸精製キットが備える弾性部材の図であり、（Ａ）は上面図、（Ｂ）は左側面図、（Ｃ）は断面図である。It is a figure of the elastic member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) is a top view, (B) is a left view, (C) is sectional drawing. 同実施形態に係る核酸精製キットが備える回転部材の図であり、（Ａ）および（Ｂ）は側面図、（Ｃ）は上面図、（Ｄ）は断面図である。It is a figure of the rotation member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) And (B) is a side view, (C) is a top view, (D) is sectional drawing. 同実施形態に係る核酸精製キットが備えるピストン部材の図であり、（Ａ）および（Ｂ）は、上下方向に垂直な方向から見た外観図である。It is a figure of the piston member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) And (B) is the external view seen from the direction perpendicular | vertical to an up-down direction. 同実施形態に係る核酸精製キットが備える磁石部材の図であり、（Ａ）は前面図、（Ｂ）は左側面図、（Ｃ）は上面図である。It is a figure of the magnetic member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) is a front view, (B) is a left view, (C) is a top view. 同実施形態に係る核酸精製キットが備える摺動制限部材の図であり、（Ａ）は上面図、（Ｂ）は断面図である。It is a figure of the sliding restriction member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) is a top view, (B) is sectional drawing. 同実施形態に係る核酸精製キットが備えるカバー部材の図であり、（Ａ）は左側面図、（Ｂ）は背面図、（Ｃ）は平面図である。It is a figure of the cover member with which the nucleic acid purification kit which concerns on the same embodiment is provided, (A) is a left view, (B) is a rear view, (C) is a top view. （Ａ）〜（Ｃ）は、同実施形態に係る回転部材の動作を説明するための断面図である。(A)-(C) are sectional drawings for demonstrating operation | movement of the rotating member which concerns on the same embodiment. （Ａ）および（Ｂ）は、同実施形態に係るピストン部材および磁石部材の動作を説明するための断面図である。(A) And (B) is sectional drawing for demonstrating operation | movement of the piston member and magnet member which concern on the embodiment. （Ａ）〜（Ｃ）は、同実施形態に係るカバー部材の動作を説明するための左側面図である。(A)-(C) are the left view for demonstrating operation | movement of the cover member which concerns on the same embodiment. 変形例に係るピストン部材の図であり、（Ａ）および（Ｂ）は、上下方向に垂直な方向から見た外観図である。It is a figure of the piston member which concerns on a modification, (A) And (B) is the external view seen from the direction perpendicular | vertical to an up-down direction. 変形例に係る摺動制限部材の図であり、（Ａ）は上面図、（Ｂ）は断面図である。It is a figure of the sliding restriction member concerning a modification, (A) is a top view and (B) is a sectional view.

図面を参照して、本発明に係る混合器具を核酸精製キット１として具現化した実施形態を説明する。なお、本実施形態の説明に含まれる上下方向、前後方向、および、左右方向は、図中の矢印で示す方向であって、互いに直交する方向である。   With reference to the drawings, an embodiment in which a mixing instrument according to the present invention is embodied as a nucleic acid purification kit 1 will be described. In addition, the up-down direction, the front-back direction, and the left-right direction included in the description of the present embodiment are directions indicated by arrows in the drawing, and are directions orthogonal to each other.

図１は、核酸精製キット１の使用前の状態を示しており、核酸精製キット１は、検体である唾液に含まれているウイルスの核酸（ＤＮＡまたはＲＮＡ）を精製するための器具本体１Ａと、精製された核酸を器具本体１Ａから排出するための排出用ノズル９０とを備えている。   FIG. 1 shows a state before use of the nucleic acid purification kit 1, and the nucleic acid purification kit 1 includes an instrument body 1A for purifying viral nucleic acid (DNA or RNA) contained in saliva as a sample. And a discharging nozzle 90 for discharging the purified nucleic acid from the instrument main body 1A.

排出用ノズル９０は、合成樹脂により形成された管である。排出用ノズル９０は、核酸精製キット１の使用時に器具本体１Ａに取り付けられる基端部９１と、基端部９１よりも尖った先端部９２とを有している。   The discharge nozzle 90 is a tube made of synthetic resin. The discharge nozzle 90 has a base end portion 91 that is attached to the instrument main body 1 </ b> A when the nucleic acid purification kit 1 is used, and a tip end portion 92 that is sharper than the base end portion 91.

図２は、図１に示した器具本体１Ａの分解斜視図である。図２に示すように、核酸精製キット１は、器具本体１Ａを構成する部品として、ベース部材１０と、弾性部材２０と、回転部材３０と、ピストン部材４０と、磁石部材５０と、摺動制限部材６０と、カバー部材７０とを備えている。   2 is an exploded perspective view of the instrument main body 1A shown in FIG. As shown in FIG. 2, the nucleic acid purification kit 1 includes a base member 10, an elastic member 20, a rotating member 30, a piston member 40, a magnet member 50, and a sliding restriction as components constituting the instrument body 1 </ b> A. A member 60 and a cover member 70 are provided.

図３を参照して、ベース部材１０について説明する。なお、図３（Ｆ）は、図３（Ａ）および（Ｂ）のＡ−Ａ線断面図である。
ベース部材１０は、ポリプロピレン樹脂等の合成樹脂により形成されている。ベース部材１０は、複数の物質を混合するための混合室１１と、回転部材３０を収容する回転部材収容部１２と、混合室１１と回転部材収容部１２との間に形成された貫通孔１３と、ベース部材１０の下端部に設けられた弾性部材保持部１４と、ベース部材１０の外表面に設けられた目印１５と、ベース部材１０の上端部に設けられた摺動制限部材係合部１６およびカバー部材支持部１７とを有している。 The base member 10 will be described with reference to FIG. Note that FIG. 3F is a cross-sectional view taken along line AA in FIGS. 3A and 3B.
The base member 10 is made of a synthetic resin such as a polypropylene resin. The base member 10 includes a mixing chamber 11 for mixing a plurality of substances, a rotating member accommodating portion 12 for accommodating the rotating member 30, and a through hole 13 formed between the mixing chamber 11 and the rotating member accommodating portion 12. An elastic member holding portion 14 provided at the lower end portion of the base member 10, a mark 15 provided on the outer surface of the base member 10, and a sliding limit member engaging portion provided at the upper end portion of the base member 10. 16 and a cover member support portion 17.

ベース部材１０の後部に設けられた混合室１１、および、ベース部材１０の前部に設けられた回転部材収容部１２は、それぞれ、上下方向に延びる貫通孔により形成され、上方および下方に開口している。前後方向に延びた貫通孔１３の一端は、混合室１１の内周面に開口し、貫通孔１３の他端は、回転部材収容部１２の一部である弾性部材保持部１４の内周面に開口している。弾性部材保持部１４は、弾性部材２０を収容する窪みにより形成されており、回転部材収容部１２の下端部を構成している。弾性部材保持部１４は、弾性部材２０を保持することで、ベース部材１０に対して弾性部材２０が動くことを規制する。目印１５は、ベース部材１０の下端部に設けられた小さな縦長の突起により形成されている。摺動制限部材係合部１６は、混合室１１の上側開口が形成された面よりも上方に突出するとともにコ字状に形成された左右一対の突起により形成されている。摺動制限部材係合部１６は、摺動制限部材６０に当接することで、ベース部材１０に対する摺動制限部材６０の前後方向および左右方向の移動を規制する。カバー部材支持部１７は、ベース部材１０の側部の外表面から左方および右方に突出するとともに円柱状に形成された左右一対の突起により形成されている。カバー部材支持部１７は、カバー部材７０を回転可能に支持する。   The mixing chamber 11 provided at the rear portion of the base member 10 and the rotating member accommodating portion 12 provided at the front portion of the base member 10 are formed by through holes extending in the vertical direction, and open upward and downward. ing. One end of the through-hole 13 extending in the front-rear direction opens to the inner peripheral surface of the mixing chamber 11, and the other end of the through-hole 13 is the inner peripheral surface of the elastic member holding portion 14 that is a part of the rotating member housing portion 12. Is open. The elastic member holding portion 14 is formed by a recess that accommodates the elastic member 20 and constitutes the lower end portion of the rotating member accommodating portion 12. The elastic member holding part 14 holds the elastic member 20 and restricts the elastic member 20 from moving with respect to the base member 10. The mark 15 is formed by a small vertically long protrusion provided at the lower end of the base member 10. The sliding restricting member engaging portion 16 is formed by a pair of left and right protrusions that protrude above the surface of the mixing chamber 11 where the upper opening is formed and that are formed in a U shape. The sliding restriction member engaging portion 16 abuts against the sliding restriction member 60 to restrict the movement of the sliding restriction member 60 relative to the base member 10 in the front-rear direction and the left-right direction. The cover member support portion 17 is formed by a pair of left and right protrusions that protrude leftward and rightward from the outer surface of the side portion of the base member 10 and are formed in a columnar shape. The cover member support part 17 supports the cover member 70 rotatably.

図４を参照して、弾性部材２０について説明する。なお、図４（Ｃ）は、図４（Ａ）のＢ−Ｂ線断面図である。
弾性部材２０は、回転部材３０を回転可能に支持する軸受部材、および、混合室１１を下方から閉塞する膜部材として機能する。弾性部材２０は、エラストマーにより形成されており、例えば、ベース部材１０の射出成形時におけるインサート成形によって弾性部材保持部１４に設けられる。弾性部材２０は、回転部材支持部２１と、膜部２２と、回転部材支持部２１に形成された貫通孔２３と、膜部２２の下方に設けられたノズル保持部２４とを有している。 The elastic member 20 will be described with reference to FIG. Note that FIG. 4C is a cross-sectional view taken along line BB in FIG.
The elastic member 20 functions as a bearing member that rotatably supports the rotating member 30 and a membrane member that closes the mixing chamber 11 from below. The elastic member 20 is formed of an elastomer, and is provided in the elastic member holding portion 14 by insert molding at the time of injection molding of the base member 10, for example. The elastic member 20 includes a rotating member support portion 21, a membrane portion 22, a through hole 23 formed in the rotating member support portion 21, and a nozzle holding portion 24 provided below the membrane portion 22. .

弾性部材２０の前部に設けられた回転部材支持部２１は、上方および下方に開口した円筒状の貫通孔により形成されている。回転部材支持部２１は、回転部材３０が圧入されることによって、回転部材３０を回転可能に支持する。弾性部材２０の後部に設けられた膜部２２は、混合室１１を下方から閉塞する。膜部２２は、排出用ノズル９０の先端部９２で穿つことが可能な厚さを有している。膜部２２を排出用ノズル９０で下方から穿つと、混合室１１内の物質を排出するための排出孔（図示略）が膜部２２に形成される。前後方向に延びた貫通孔２３の一端は、回転部材支持部２１の内周面に開口し、貫通孔２３の他端は、弾性部材２０の外表面に開口している。貫通孔２３は、ベース部材１０の貫通孔１３と連なる。ノズル保持部２４は、排出用ノズル９０の基端部９１が下方から圧入される窪みにより形成されている。ノズル保持部２４は、基端部９１を保持することで、排出用ノズル９０が排出孔に取り付けられた状態を維持する。   The rotating member support portion 21 provided at the front portion of the elastic member 20 is formed by a cylindrical through hole that opens upward and downward. The rotating member support portion 21 supports the rotating member 30 so that the rotating member 30 can be rotated when the rotating member 30 is press-fitted. The film part 22 provided at the rear part of the elastic member 20 closes the mixing chamber 11 from below. The film portion 22 has a thickness that can be pierced by the tip end portion 92 of the discharge nozzle 90. When the film part 22 is pierced from below by the discharge nozzle 90, a discharge hole (not shown) for discharging the substance in the mixing chamber 11 is formed in the film part 22. One end of the through hole 23 extending in the front-rear direction opens to the inner peripheral surface of the rotating member support portion 21, and the other end of the through hole 23 opens to the outer surface of the elastic member 20. The through hole 23 is continuous with the through hole 13 of the base member 10. The nozzle holding portion 24 is formed by a recess into which the base end portion 91 of the discharge nozzle 90 is press-fitted from below. The nozzle holding unit 24 maintains the state where the discharge nozzle 90 is attached to the discharge hole by holding the base end portion 91.

図５を参照して、回転部材３０について説明する。なお、図５（Ｃ）において、図５（Ａ）および（Ｂ）に示す蓋３０Ｂの図示は省略されている。また、図５（Ｄ）は、図５（Ａ）および（Ｂ）のＣ−Ｃ線断面図である。
回転部材３０は、弾性部材２０に圧入されることで、ベース部材１０および弾性部材２０に対して、上下方向に平行な軸線を中心として回転可能に設けられる。回転部材３０は、混合する物質を貯留する貯留器３０Ａと、貯留器３０Ａの上端部に設けられる蓋３０Ｂとにより構成されている。 The rotating member 30 will be described with reference to FIG. In FIG. 5C, illustration of the lid 30B shown in FIGS. 5A and 5B is omitted. FIG. 5D is a cross-sectional view taken along line CC of FIGS. 5A and 5B.
The rotating member 30 is press-fitted into the elastic member 20 so as to be rotatable with respect to the base member 10 and the elastic member 20 around an axis parallel to the vertical direction. The rotating member 30 includes a reservoir 30A that stores a substance to be mixed, and a lid 30B that is provided at the upper end of the reservoir 30A.

貯留器３０Ａは、ベース部材１０と同様に合成樹脂により形成されている。貯留器３０Ａは、基板部３１と、基板部３１から上方に立ち上がっている円柱部３２と、混合する物質を貯留するための複数の貯留室３３Ａ〜３３Ｃと、弾性部材２０に圧入される圧入部３４と、圧入部３４に設けられた貫通孔３５Ａ〜３５Ｃとを有している。   The reservoir 30 </ b> A is formed of a synthetic resin like the base member 10. The reservoir 30 </ b> A includes a substrate portion 31, a cylindrical portion 32 rising upward from the substrate portion 31, a plurality of storage chambers 33 </ b> A to 33 </ b> C for storing a substance to be mixed, and a press-fit portion that is press-fitted into the elastic member 20. 34 and through holes 35 </ b> A to 35 </ b> C provided in the press-fit portion 34.

ベース部材１０の下方に設けられた基板部３１は、円盤状に形成されている。円柱部３２は、ベース部材１０の回転部材収容部１２に挿し込まれて収容される。貯留室３３Ａ〜３３Ｃは、それぞれ、円柱部３２の内部に設けられ、上方に開口した有底孔により形成されている。貯留室３３Ａ〜３３Ｃの容積は異なっており、貯留室３３Ａの容積は貯留室３３Ｃの容積よりも大きく、かつ、貯留室３３Ｂの容積は貯留室３３Ｃの容積よりも小さい。圧入部３４は、円柱部３２の下端部を構成しており、弾性部材２０の回転部材支持部２１に圧入される。貫通孔３５Ａの一端は、貯留室３３Ａの底部に開口し、貫通孔３５Ａの他端は、圧入部３４の外周面に開口している。貫通孔３５Ｂの一端は、貯留室３３Ｂの底部に開口し、貫通孔３５Ｂの他端は、圧入部３４の外周面に開口している。貫通孔３５Ｃの一端は、貯留室３３Ｃの底部に開口し、貫通孔３５Ｃの他端は、圧入部３４の外周面に開口している。圧入部３４の外周面における貫通孔３５Ａ〜３５Ｃの開口は、上方から見て円柱部３２の周方向において間隔をあけて設けられており、かつ、上下方向において同じ高さに位置している。   The substrate portion 31 provided below the base member 10 is formed in a disk shape. The cylindrical part 32 is inserted and accommodated in the rotating member accommodating part 12 of the base member 10. Each of the storage chambers 33A to 33C is formed by a bottomed hole that is provided inside the cylindrical portion 32 and opens upward. The volumes of the storage chambers 33A to 33C are different, the volume of the storage chamber 33A is larger than the volume of the storage chamber 33C, and the volume of the storage chamber 33B is smaller than the volume of the storage chamber 33C. The press-fit portion 34 constitutes the lower end portion of the cylindrical portion 32 and is press-fitted into the rotating member support portion 21 of the elastic member 20. One end of the through hole 35A opens to the bottom of the storage chamber 33A, and the other end of the through hole 35A opens to the outer peripheral surface of the press-fit portion 34. One end of the through hole 35B opens to the bottom of the storage chamber 33B, and the other end of the through hole 35B opens to the outer peripheral surface of the press-fit portion 34. One end of the through hole 35C opens to the bottom of the storage chamber 33C, and the other end of the through hole 35C opens to the outer peripheral surface of the press-fit portion 34. The openings of the through holes 35 </ b> A to 35 </ b> C on the outer peripheral surface of the press-fit portion 34 are provided at intervals in the circumferential direction of the columnar portion 32 when viewed from above, and are positioned at the same height in the vertical direction.

また、貯留器３０Ａは、円柱部３２の外周面に沿って設けられた環状溝３６と、基板部３１の外周面に設けられた目印３７Ａ〜３７Ｃと、基板部３１から側方に突出する一対のフランジ３８とを有している。   The reservoir 30 </ b> A includes a ring groove 36 provided along the outer peripheral surface of the cylindrical portion 32, marks 37 </ b> A to 37 </ b> C provided on the outer peripheral surface of the substrate portion 31, and a pair protruding laterally from the substrate portion 31. The flange 38 is provided.

環状溝３６は、回転部材収容部１２よりも上方に突出する円柱部３２の上端部に形成されている。目印３７Ａ〜３７Ｃは、それぞれ、基板部３１の外表面に設けられた小さな縦長の突起により形成されている。目印３７Ａは、貫通孔３５Ａ，２３が連なったときに目印１５と上下に並ぶ位置に設けられている。目印３７Ｂは、貫通孔３５Ｂ，２３が連なったときに目印１５と上下に並ぶ位置に設けられている。目印３７Ｃは、貫通孔３５Ｃ，２３が連なったときに目印１５と上下に並ぶ位置に設けられている。フランジ３８は、回転部材３０を回転させる使用者が持ちやすい形状に形成されている。   The annular groove 36 is formed at the upper end portion of the cylindrical portion 32 that protrudes upward from the rotating member housing portion 12. Each of the marks 37 </ b> A to 37 </ b> C is formed by a small vertically long protrusion provided on the outer surface of the substrate portion 31. The mark 37A is provided at a position aligned vertically with the mark 15 when the through holes 35A and 23 are connected. The mark 37 </ b> B is provided at a position aligned vertically with the mark 15 when the through holes 35 </ b> B and 23 are connected. The mark 37C is provided at a position aligned vertically with the mark 15 when the through holes 35C and 23 are connected. The flange 38 is formed in a shape that can be easily held by a user who rotates the rotating member 30.

蓋３０Ｂは、エラストマーにより形成されており、貯留器３０Ａに対して取り外し可能に構成されている。蓋３０Ｂは、貯留室３３Ａ〜３３Ｃの上側開口に圧入されることで、貯留室３３Ａ〜３３Ｃを上方から閉塞する。   The lid 30B is made of an elastomer and is configured to be removable from the reservoir 30A. The lid 30B closes the storage chambers 33A to 33C from above by being press-fitted into the upper openings of the storage chambers 33A to 33C.

図６を参照して、ピストン部材４０について説明する。なお、図６（Ａ）および（Ｂ）は、上下方向に垂直な方向から見たピストン部材４０の外観図である。
ピストン部材４０は、ベース部材１０の混合室１１に挿し込まれることで、ベース部材１０に対して上下方向に摺動可能に設けられる。ピストン部材４０の摺動量（上下方向の移動量）と、混合室１１内の容積の変化量とは、比例の関係にある。ピストン部材４０は、混合室１１内に設けられるピストンヘッド４１と、ピストンヘッド４１が下端に取り付けられたピストンロッド４２と、ピストンロッド４２の上端に設けられたリング４３と、ピストンロッド４２の側部に設けられた突部４４，４５とを有している。 The piston member 40 will be described with reference to FIG. 6A and 6B are external views of the piston member 40 as viewed from a direction perpendicular to the vertical direction.
The piston member 40 is slidable in the vertical direction with respect to the base member 10 by being inserted into the mixing chamber 11 of the base member 10. The sliding amount (vertical movement amount) of the piston member 40 and the amount of change in the volume in the mixing chamber 11 are in a proportional relationship. The piston member 40 includes a piston head 41 provided in the mixing chamber 11, a piston rod 42 to which the piston head 41 is attached at the lower end, a ring 43 provided at the upper end of the piston rod 42, and a side portion of the piston rod 42. And protrusions 44 and 45 provided on the surface.

ピストンヘッド４１は、エラストマーにより形成されたガスケットである。ピストンヘッド４１は、混合室１１を上方から閉塞し、混合室１１の内周面に当接した状態で、ベース部材１０に対して上下方向に移動する。合成樹脂により形成されたピストンロッド４２は、ピストンヘッド４１とリング４３とを接続しており、混合室１１の長さ方向である上下方向に延びる。ピストンロッド４２は、混合室１１の上側開口から上方に突出するように設けられ、ベース部材１０の内部から外部に延びる。リング４３および突部４４，４５は、合成樹脂によりピストンロッド４２と一体として形成されている。リング４３は、指を入れることができる程度の大きさを有しており、ベース部材１０よりも上方に配置される。突部４４は、ピストンロッド４２が延びる上下方向において所定間隔をあけて複数設けられ、上下方向に対して斜めに延びている。突部４５は、上下方向に直線状に設けられ、各突部４４の一端部と一体として形成されている。   The piston head 41 is a gasket formed of an elastomer. The piston head 41 closes the mixing chamber 11 from above and moves in the vertical direction with respect to the base member 10 in a state where the mixing chamber 11 is in contact with the inner peripheral surface of the mixing chamber 11. The piston rod 42 made of synthetic resin connects the piston head 41 and the ring 43 and extends in the vertical direction, which is the length direction of the mixing chamber 11. The piston rod 42 is provided so as to protrude upward from the upper opening of the mixing chamber 11 and extends from the inside of the base member 10 to the outside. The ring 43 and the protrusions 44 and 45 are formed integrally with the piston rod 42 from a synthetic resin. The ring 43 has a size that allows a finger to be inserted, and is disposed above the base member 10. A plurality of protrusions 44 are provided at predetermined intervals in the vertical direction in which the piston rod 42 extends, and extend obliquely with respect to the vertical direction. The protrusion 45 is provided in a straight line in the vertical direction, and is formed integrally with one end of each protrusion 44.

図７を参照して、磁石部材５０について説明する。
磁石部材５０は、混合室１１の内部に磁場を発生させることで、混合室１１内の磁性体を磁力で吸引する。磁石部材５０は、ベース部材１０の後端部に取り付けられることで、ベース部材１０に対して上下方向に摺動可能に設けられる。磁石部材５０は、磁場を発生させる永久磁石５１と、永久磁石５１を保持する磁石ホルダー５２と、ベース部材１０を保持するベース部材保持部５３とを有している。 The magnet member 50 will be described with reference to FIG.
The magnet member 50 attracts the magnetic material in the mixing chamber 11 with a magnetic force by generating a magnetic field inside the mixing chamber 11. The magnet member 50 is provided to be slidable in the vertical direction with respect to the base member 10 by being attached to the rear end portion of the base member 10. The magnet member 50 includes a permanent magnet 51 that generates a magnetic field, a magnet holder 52 that holds the permanent magnet 51, and a base member holding portion 53 that holds the base member 10.

硬質磁性体により形成された永久磁石５１は、磁石ホルダー５２に埋め込まれている。合成樹脂により形成された磁石ホルダー５２は、永久磁石５１を保持している。ベース部材保持部５３は、合成樹脂により磁石ホルダー５２と一体として形成されるとともに、永久磁石５１よりも前方に突出した左右一対の突起により形成されている。ベース部材保持部５３は、ベース部材１０の後端部を挟むことによって、ベース部材１０に磁石部材５０が取り付けられた状態を維持する。   A permanent magnet 51 formed of a hard magnetic material is embedded in a magnet holder 52. A magnet holder 52 formed of synthetic resin holds the permanent magnet 51. The base member holding portion 53 is formed integrally with the magnet holder 52 from a synthetic resin, and is formed by a pair of left and right protrusions that protrude forward from the permanent magnet 51. The base member holding portion 53 maintains the state where the magnet member 50 is attached to the base member 10 by sandwiching the rear end portion of the base member 10.

図８を参照して、摺動制限部材６０について説明する。なお、図８（Ｂ）は、図８（Ａ）のＤ−Ｄ線断面図である。
摺動制限部材６０は、合成樹脂により形成されており、左右一対の摺動制限部材係合部１６の間においてベース部材１０の上面に載置されることで、ベース部材１０に対して前後方向に移動可能に設けられる。摺動制限部材６０は、ピストン部材４０の突部４４，４５に当接することで、上下方向におけるピストン部材４０の摺動を制限する。摺動制限部材６０は、ピストン部材４０の摺動を制限することによって、混合室１１内の容積の変化を制限し、混合室１１外への物質の吐出量を制限する。摺動制限部材６０は、ピストンロッド４２の後方に設けられる基部６１と、基部６１から前方に突出した左右一対の突出部６２と、基部６１に設けられたピストン部材係合部６３と、突出部６２に設けられたベース部材係合部６４とを有している。 The sliding restriction member 60 will be described with reference to FIG. Note that FIG. 8B is a cross-sectional view taken along the line DD in FIG.
The sliding restriction member 60 is made of synthetic resin, and is placed on the upper surface of the base member 10 between the pair of left and right sliding restriction member engaging portions 16, so that the longitudinal direction with respect to the base member 10. It is provided to be movable. The sliding limit member 60 limits the sliding of the piston member 40 in the vertical direction by contacting the protrusions 44 and 45 of the piston member 40. The sliding restriction member 60 restricts the volume change in the mixing chamber 11 by restricting the sliding of the piston member 40, and restricts the discharge amount of the substance to the outside of the mixing chamber 11. The sliding restriction member 60 includes a base 61 provided behind the piston rod 42, a pair of left and right protrusions 62 protruding forward from the base 61, a piston member engaging part 63 provided on the base 61, and a protrusion. And a base member engaging portion 64 provided at 62.

基部６１は、ベース部材１０上において混合室１１の上側開口よりも後方に載置され、左右一対の突出部６２は、ベース部材１０上において混合室１１の上側開口を挟むように配置される。ピストン部材係合部６３は、基部６１から前方に突出した突起により形成されている。ピストン部材係合部６３は、ピストン部材４０の複数の突部４４の間に挿し込まれ、突部４４，４５に当接することによって、ピストン部材４０の上下方向の移動を規制する。ベース部材係合部６４は、各突出部６２の側部から左右方向に突出した左右一対の突起により形成されている。ベース部材係合部６４が、ベース部材１０の摺動制限部材係合部１６に当接することによって、摺動制限部材６０の前後方向の移動が規制される。   The base 61 is placed behind the upper opening of the mixing chamber 11 on the base member 10, and the pair of left and right protrusions 62 are arranged on the base member 10 so as to sandwich the upper opening of the mixing chamber 11. The piston member engaging portion 63 is formed by a protrusion protruding forward from the base portion 61. The piston member engaging portion 63 is inserted between the plurality of protrusions 44 of the piston member 40 and abuts against the protrusions 44 and 45, thereby restricting the movement of the piston member 40 in the vertical direction. The base member engaging portion 64 is formed by a pair of left and right protrusions that protrude in the left-right direction from the sides of each protrusion 62. When the base member engaging portion 64 contacts the sliding limit member engaging portion 16 of the base member 10, the movement of the sliding limit member 60 in the front-rear direction is restricted.

図９を参照して、カバー部材７０について説明する。
カバー部材７０は、合成樹脂により形成されており、回転部材３０の上端部を覆うことが可能な形状、かつ、回転部材３０の上端部を覆っている状態においてベース部材１０の前面および側面と面一になる形状を有している。カバー部材７０は、ベース部材１０に対して前後方向に移動可能に設けられるとともに、ベース部材１０に対して左右方向に平行な軸線を中心として回転可能に設けられる。カバー部材７０は、ベース部材１０のカバー部材支持部１７により支持される被支持部７１と、回転部材３０の環状溝３６に嵌まる回転部材係合部７２とを有している。 The cover member 70 will be described with reference to FIG.
The cover member 70 is formed of synthetic resin, has a shape that can cover the upper end portion of the rotating member 30, and the front surface, side surface, and surface of the base member 10 in a state that covers the upper end portion of the rotating member 30. It has a shape that becomes one. The cover member 70 is provided so as to be movable in the front-rear direction with respect to the base member 10, and is provided so as to be rotatable about an axis parallel to the left-right direction with respect to the base member 10. The cover member 70 includes a supported portion 71 supported by the cover member support portion 17 of the base member 10 and a rotating member engaging portion 72 that fits in the annular groove 36 of the rotating member 30.

被支持部７１は、カバー部材７０の後端部に設けられ、長円形状に形成された左右一対の窪みにより形成されている。回転部材係合部７２は、カバー部材７０の下端部に設けられ、カバー部材７０の内面から回転部材３０に向けて突出する突起により形成されている。回転部材係合部７２は、回転部材３０の環状溝３６に嵌まることで、ベース部材１０に対して回転部材３０が上下方向に移動することを規制する。また、回転部材係合部７２が環状溝３６に嵌まることで、カバー部材７０の回転が規制される。   The supported portion 71 is provided at the rear end portion of the cover member 70 and is formed by a pair of left and right recesses formed in an oval shape. The rotating member engaging portion 72 is provided at the lower end portion of the cover member 70, and is formed by a protrusion that protrudes from the inner surface of the cover member 70 toward the rotating member 30. The rotation member engaging portion 72 is fitted into the annular groove 36 of the rotation member 30, thereby restricting the rotation member 30 from moving in the vertical direction with respect to the base member 10. Further, the rotation of the cover member 70 is restricted by the rotation member engaging portion 72 being fitted in the annular groove 36.

次に、図１０を参照して、回転部材３０の動作について説明する。なお、図１０で示す断面は、図５（Ａ）および（Ｂ）のＣ−Ｃ線と同じ位置におけるベース部材１０および弾性部材２０の断面である。また、図１０では、蓋３０Ｂの図示を省略して回転部材３０を図示するとともに、磁石部材５０の図示を省略している。   Next, the operation of the rotating member 30 will be described with reference to FIG. In addition, the cross section shown in FIG. 10 is a cross section of the base member 10 and the elastic member 20 in the same position as the CC line | wire of FIG. 5 (A) and (B). In FIG. 10, the illustration of the lid 30 </ b> B is omitted, the rotary member 30 is shown, and the magnet member 50 is not shown.

図１０（Ａ）は、貯留室３３Ａと混合室１１とが連通している連通状態を示す断面図である。貯留室３３Ａと混合室１１とは、貫通孔３５Ａ，２３，１３を介して連通する。貯留室３３Ａと混合室１１とが連通しているときは、回転部材３０の目印３７Ａ（図５参照）が目印１５の真下に位置する。また、貯留室３３Ａと混合室１１とが連通しているときは、貫通孔３５Ｂ，３５Ｃは弾性部材２０によって覆われるため、貯留室３３Ｂ，３３Ｃは混合室１１と連通せずに弾性部材２０と蓋３０Ｂとで密閉される。すなわち、貯留室３３Ａと混合室１１との連通状態においては、貯留室３３Ｂ，３３Ｃと混合室１１とは連通していない非連通状態である。   FIG. 10A is a cross-sectional view showing a communication state in which the storage chamber 33A and the mixing chamber 11 communicate with each other. The storage chamber 33A and the mixing chamber 11 communicate with each other through the through holes 35A, 23, and 13. When the storage chamber 33 </ b> A and the mixing chamber 11 communicate with each other, the mark 37 </ b> A (see FIG. 5) of the rotating member 30 is located directly below the mark 15. Further, when the storage chamber 33A and the mixing chamber 11 are in communication with each other, the through holes 35B and 35C are covered with the elastic member 20, so that the storage chambers 33B and 33C are not in communication with the mixing chamber 11 and the elastic member 20 is connected. Sealed with the lid 30B. That is, in the communication state between the storage chamber 33A and the mixing chamber 11, the storage chambers 33B and 33C and the mixing chamber 11 are not in communication with each other.

図１０（Ｂ）は、貯留室３３Ｂと混合室１１とが連通している連通状態を示す断面図である。貯留室３３Ｂと混合室１１とは、貫通孔３５Ｂ，２３，１３を介して連通する。貯留室３３Ｂと混合室１１とが連通しているときは、回転部材３０の目印３７Ｂ（図５参照）が目印１５の真下に位置する。また、貯留室３３Ｂと混合室１１とが連通しているときは、貫通孔３５Ａ，３５Ｃは弾性部材２０によって覆われるため、貯留室３３Ａ，３３Ｃは混合室１１と連通せずに弾性部材２０と蓋３０Ｂとで密閉される。すなわち、貯留室３３Ｂと混合室１１との連通状態においては、貯留室３３Ａ，３３Ｃと混合室１１とは連通していない非連通状態である。   FIG. 10B is a cross-sectional view showing a communication state in which the storage chamber 33B and the mixing chamber 11 communicate with each other. The storage chamber 33B and the mixing chamber 11 communicate with each other through the through holes 35B, 23, and 13. When the storage chamber 33 </ b> B and the mixing chamber 11 communicate with each other, the mark 37 </ b> B (see FIG. 5) of the rotating member 30 is located directly below the mark 15. Further, when the storage chamber 33B and the mixing chamber 11 are in communication, the through holes 35A and 35C are covered with the elastic member 20, so that the storage chambers 33A and 33C are not in communication with the mixing chamber 11 and the elastic member 20 is connected. Sealed with the lid 30B. That is, in the communication state between the storage chamber 33B and the mixing chamber 11, the storage chambers 33A and 33C and the mixing chamber 11 are not in communication with each other.

図１０（Ｃ）は、貯留室３３Ｃと混合室１１とが連通している連通状態を示す断面図である。貯留室３３Ｃと混合室１１とは、貫通孔３５Ｃ，２３，１３を介して連通する。貯留室３３Ｃと混合室１１とが連通しているときは、回転部材３０の目印３７Ｃ（図５参照）が目印１５の真下に位置する。また、貯留室３３Ｃと混合室１１とが連通しているときは、貫通孔３５Ａ，３５Ｂは弾性部材２０によって覆われるため、貯留室３３Ａ，３３Ｂは混合室１１と連通せずに弾性部材２０と蓋３０Ｂとで密閉される。すなわち、貯留室３３Ｃと混合室１１との連通状態においては、貯留室３３Ａ，３３Ｂと混合室１１とは連通していない非連通状態である。   FIG. 10C is a cross-sectional view showing a communication state in which the storage chamber 33C and the mixing chamber 11 communicate with each other. The storage chamber 33C and the mixing chamber 11 communicate with each other through the through holes 35C, 23, and 13. When the storage chamber 33 </ b> C and the mixing chamber 11 communicate with each other, the mark 37 </ b> C (see FIG. 5) of the rotating member 30 is located directly below the mark 15. Further, when the storage chamber 33C and the mixing chamber 11 communicate with each other, the through holes 35A and 35B are covered with the elastic member 20, so that the storage chambers 33A and 33B do not communicate with the mixing chamber 11 and the elastic member 20 does not communicate with each other. Sealed with the lid 30B. That is, in the communication state between the storage chamber 33C and the mixing chamber 11, the storage chambers 33A and 33B and the mixing chamber 11 are not in communication with each other.

図１０（Ａ）に示す回転部材３０を例えば時計回り方向に１５０°回転することによって、貯留室３３Ｂと混合室１１との連通状態、すなわち図１０（Ｂ）に示す状態になる。また、図１０（Ｂ）に示す回転部材３０を例えば時計回り方向に６０°回転することによって、貯留室３３Ｃと混合室１１との連通状態、すなわち図１０（Ｃ）に示す状態になる。このように、貯留室３３Ａ〜３３Ｃの全てが混合室１１と同時に連通することはなく、回転部材３０の回転角が特定の角度であるとき、貯留室３３Ａ〜３３Ｃのうち１つと混合室１１とが連通する連通状態となり、他の２つと混合室１１とが連通していない非連通状態となる。そして、ベース部材１０に対して回転部材３０が回転することで、回転部材３０の回転角が変化し、貯留室３３Ａ〜３３Ｃと混合室１１との連通状態および非連通状態が切り替わる。また、貫通孔３５Ａ〜３５Ｃが弾性部材２０によって覆われるように回転部材３０の回転角を維持した場合には、混合室１１が貯留室３３Ａ〜３３Ｃのいずれとも連通せず、混合室１１および貯留室３３Ａ〜３３Ｃの各々が密閉された状態となる。   By rotating the rotating member 30 shown in FIG. 10A, for example, by 150 ° in the clockwise direction, the communication state between the storage chamber 33B and the mixing chamber 11, that is, the state shown in FIG. Further, by rotating the rotating member 30 shown in FIG. 10B by, for example, 60 ° in the clockwise direction, the communication state between the storage chamber 33C and the mixing chamber 11, that is, the state shown in FIG. Thus, not all of the storage chambers 33A to 33C communicate with the mixing chamber 11 at the same time, and when the rotation angle of the rotating member 30 is a specific angle, one of the storage chambers 33A to 33C and the mixing chamber 11 Is in a communication state in which the other two are not in communication with the mixing chamber 11. And the rotation angle of the rotation member 30 changes because the rotation member 30 rotates with respect to the base member 10, and the communication state and the non-communication state of the storage chambers 33A-33C and the mixing chamber 11 are switched. When the rotation angle of the rotating member 30 is maintained so that the through holes 35A to 35C are covered with the elastic member 20, the mixing chamber 11 does not communicate with any of the storage chambers 33A to 33C, and the mixing chamber 11 and the storage chamber 11 are stored. Each of the chambers 33A to 33C is in a sealed state.

図１１を参照して、ピストン部材４０と磁石部材５０の動作について説明する。なお、ピストン部材４０の動作と磁石部材５０の動作は、それぞれ独立した動作である。また、図１１で示す断面は、図３（Ａ）および（Ｂ）のＡ−Ａ線と同じ位置におけるベース部材１０、弾性部材２０、磁石部材５０、摺動制限部材６０、および、カバー部材７０の断面である。   The operation of the piston member 40 and the magnet member 50 will be described with reference to FIG. The operation of the piston member 40 and the operation of the magnet member 50 are independent operations. Moreover, the cross section shown in FIG. 11 is the base member 10, the elastic member 20, the magnet member 50, the sliding limiting member 60, and the cover member 70 in the same position as the AA line of FIG. 3 (A) and (B). It is a cross section.

ピストン部材４０は、図１１（Ａ）に示すリング４３を持ってピストンロッド４２を持ち上げることによって、図１１（Ｂ）に示すように、ピストンヘッド４１が混合室１１の内周面に対して当接した状態で上方に移動する。こうして、ピストン部材４０を上方に摺動させると、混合室１１内の容積が拡大する。貯留室３３Ａ〜３３Ｃのいずれか１つと混合室１１とが連通しているときに混合室１１内の容積が拡大すると、混合室１１と連通状態になっている貯留室３３Ａ〜３３Ｃのいずれか（以下、「連通貯留室」という）が貯留している物質が、連通貯留室から混合室１１に吸入される。   The piston member 40 holds the ring 43 shown in FIG. 11 (A) and lifts the piston rod 42 so that the piston head 41 contacts the inner peripheral surface of the mixing chamber 11 as shown in FIG. 11 (B). Moves up in contact. Thus, when the piston member 40 is slid upward, the volume in the mixing chamber 11 increases. If the volume in the mixing chamber 11 is expanded when any one of the storage chambers 33A to 33C is in communication with the mixing chamber 11, any of the storage chambers 33A to 33C in communication with the mixing chamber 11 ( Hereinafter, the substance stored in the “communication storage chamber” is sucked into the mixing chamber 11 from the communication storage chamber.

また、ピストン部材４０は、図１１（Ｂ）に示すリング４３を持ってピストンロッド４２を押し下げることによって、図１１（Ａ）に示すように、ピストンヘッド４１が混合室１１の内周面に対して当接した状態で下方に移動する。こうして、ピストン部材４０を下方に摺動させると、混合室１１内の容積が縮小する。貯留室３３Ａ〜３３Ｃのいずれか１つと混合室１１とが連通しているときに混合室１１内の容積が縮小すると、混合室１１が貯留している物質が、混合室１１から連通貯留室に吐出される。また、膜部２２に排出孔（図示略）が形成されている状態で混合室１１内の容積が縮小すると、混合室１１が貯留している物質が、膜部２２の排出孔から排出される。   Further, the piston member 40 holds the ring 43 shown in FIG. 11 (B) and pushes down the piston rod 42, so that the piston head 41 moves against the inner peripheral surface of the mixing chamber 11 as shown in FIG. 11 (A). And move downward in contact. Thus, when the piston member 40 is slid downward, the volume in the mixing chamber 11 is reduced. When the volume in the mixing chamber 11 is reduced when any one of the storage chambers 33A to 33C is in communication with the mixing chamber 11, the substance stored in the mixing chamber 11 is transferred from the mixing chamber 11 to the communication storage chamber. Discharged. Further, when the volume in the mixing chamber 11 is reduced in a state where the discharge hole (not shown) is formed in the film part 22, the substance stored in the mixing chamber 11 is discharged from the discharge hole of the film part 22. .

なお、図１１（Ａ）に示すピストン部材４０は、複数の突部４４の間に摺動制限部材６０のピストン部材係合部６３が挿し込まれることによって、上下方向における摺動が規制されている。ピストン部材４０を上下方向に摺動させるためには、図１１（Ｂ）に示すように、複数の突部４４の間からピストン部材係合部６３が抜け出るように摺動制限部材６０を後方に移動させて、ピストン部材４０の摺動の制限を解除する。摺動制限部材６０の後方への移動は、ピストン部材４０を捻ってピストンロッド４２で摺動制限部材６０を押すことにより可能である。   Note that the piston member 40 shown in FIG. 11A is restricted from sliding in the vertical direction by inserting the piston member engaging portion 63 of the sliding limiting member 60 between the plurality of protrusions 44. Yes. In order to slide the piston member 40 in the vertical direction, as shown in FIG. 11B, the sliding restriction member 60 is moved backward so that the piston member engaging portion 63 comes out from between the plurality of protrusions 44. It is moved and the restriction | limiting of the sliding of the piston member 40 is cancelled | released. The rearward movement of the sliding restriction member 60 is possible by twisting the piston member 40 and pushing the sliding restriction member 60 with the piston rod 42.

図１１（Ａ）および（Ｂ）に示す磁石部材５０は、ベース部材１０に密着した状態を維持しながら、上下方向に移動することが可能である。磁石部材５０は、混合室１１内の磁石部材５０近傍の磁性体を吸引することで、混合室１１内に磁性体を凝集させる。図１１（Ａ）に示すように混合室１１の下端部近傍に位置する磁石部材５０は、貫通孔１３近傍で磁性体を吸引する。磁石部材５０は、上下方向に往復移動することによって、混合室１１の全体に分散した磁性体を吸引することが可能である。また、図１１（Ａ）に示す磁石部材５０を上方に移動させると、ベース部材１０から簡単に取り外すことが可能である。ピストンヘッド４１よりも上方に磁石部材５０を配置したり、ベース部材１０から磁石部材５０を取り外したりすることで、混合室１１内の磁性体は吸引されない。   The magnet member 50 shown in FIGS. 11A and 11B can move in the vertical direction while maintaining a state of being in close contact with the base member 10. The magnet member 50 attracts the magnetic body in the vicinity of the magnet member 50 in the mixing chamber 11 to aggregate the magnetic body in the mixing chamber 11. As shown in FIG. 11A, the magnet member 50 located near the lower end of the mixing chamber 11 attracts the magnetic material in the vicinity of the through hole 13. The magnet member 50 is capable of attracting the magnetic material dispersed throughout the mixing chamber 11 by reciprocating in the vertical direction. Further, when the magnet member 50 shown in FIG. 11A is moved upward, it can be easily detached from the base member 10. By arranging the magnet member 50 above the piston head 41 or removing the magnet member 50 from the base member 10, the magnetic body in the mixing chamber 11 is not attracted.

図１２を参照して、カバー部材７０の動作について説明する。なお、図１２（Ａ）〜（Ｃ）は、カバー部材７０が閉状態から開状態に遷移する工程を示している。図１２（Ａ）に示すように、閉状態のカバー部材７０は、回転部材３０の上端部を覆うことで、回転部材３０の上方の空間を閉鎖している。また、図１２（Ｃ）に示すように、開状態のカバー部材７０は、回転部材３０の上方の空間を開放している。   The operation of the cover member 70 will be described with reference to FIG. 12A to 12C show a process in which the cover member 70 transitions from the closed state to the open state. As shown in FIG. 12A, the closed cover member 70 covers the upper end portion of the rotating member 30 to close the space above the rotating member 30. Further, as shown in FIG. 12C, the open cover member 70 opens a space above the rotating member 30.

図１２（Ａ）に示すカバー部材７０は、回転部材３０の環状溝３６（図５参照）に回転部材係合部７２が嵌まることで、カバー部材支持部１７を中心とする回転が規制されている。したがって、カバー部材７０を回転させるためには、図１２（Ａ）に示すカバー部材７０をベース部材１０に対して前方に移動させることで、環状溝３６から回転部材係合部７２を外す。また、左右方向に延びたリング４３がカバー部材７０の回転を規制しないように、図１２（Ａ）に示すピストン部材４０を回転させることで、図１２（Ｂ）に示すようにリング４３を前後方向に延ばす。   The cover member 70 shown in FIG. 12A is restricted from rotating around the cover member support portion 17 by fitting the rotation member engagement portion 72 into the annular groove 36 (see FIG. 5) of the rotation member 30. ing. Therefore, in order to rotate the cover member 70, the rotation member engaging portion 72 is removed from the annular groove 36 by moving the cover member 70 shown in FIG. Further, by rotating the piston member 40 shown in FIG. 12A so that the ring 43 extending in the left-right direction does not restrict the rotation of the cover member 70, the ring 43 is moved back and forth as shown in FIG. Extend in the direction.

図１２（Ｂ）は、環状溝３６から回転部材係合部７２が外れ、かつ、リング４３が前後方向に延びている状態を示している。すなわち、図１２（Ｂ）に示すカバー部材７０は、左右方向に平行な軸となるカバー部材支持部１７を中心に回転可能な状態である。図１２（Ｂ）に示すカバー部材７０は、カバー部材支持部１７を中心に回転することで、図１２（Ｃ）に示す開状態になる。図１２（Ｃ）においては、回転部材３０の上端部が露出しており、貯留器３０Ａから蓋３０Ｂを取り外して貯留室３３Ａ（図５参照）に検体を入れることが可能である。   FIG. 12B shows a state where the rotating member engaging portion 72 is detached from the annular groove 36 and the ring 43 extends in the front-rear direction. That is, the cover member 70 shown in FIG. 12B is in a state of being rotatable around the cover member support portion 17 serving as an axis parallel to the left-right direction. The cover member 70 shown in FIG. 12B is turned to the open state shown in FIG. 12C by rotating around the cover member support portion 17. In FIG. 12C, the upper end portion of the rotating member 30 is exposed, and the specimen can be put into the storage chamber 33A (see FIG. 5) by removing the lid 30B from the storage device 30A.

以上のようにして、貯留室３３Ａに検体を入れる際には、カバー部材７０を回転させて、カバー部材７０を閉状態から開状態に遷移させる。また、貯留室３３Ａに検体を入れた後は、上記手順の逆の手順で、カバー部材７０を開状態から閉状態に遷移させる。   As described above, when the specimen is put into the storage chamber 33A, the cover member 70 is rotated to change the cover member 70 from the closed state to the open state. Moreover, after putting the sample into the storage chamber 33A, the cover member 70 is changed from the open state to the closed state in the reverse order of the above procedure.

次に、核酸精製キット１を用いた核酸精製方法について説明する。なお、核酸精製方法の実施においては、貯留室３３Ａが糖鎖固定化磁性粒子を貯留し、貯留室３３Ｂが殺ウイルス剤を貯留し、貯留室３３Ｃが希釈用の水を貯留し、混合室１１が磁性粒子を貯留し、貯留室３３Ａ〜３３Ｃと混合室１１とが非連通状態であることを前提とする。   Next, a nucleic acid purification method using the nucleic acid purification kit 1 will be described. In the implementation of the nucleic acid purification method, the storage chamber 33A stores sugar chain-immobilized magnetic particles, the storage chamber 33B stores a virusicide, the storage chamber 33C stores dilution water, and the mixing chamber 11 Is assumed to store magnetic particles, and the storage chambers 33A to 33C and the mixing chamber 11 are in a non-communication state.

貯留室３３Ａが貯留する糖鎖固定化磁性粒子は、磁性を有するとともに糖鎖が固定化された金属粒子である。具体的には、糖鎖固定化磁性粒子は、糖鎖およびリンカー化合物で構成されたリガンド複合体と、金属ナノ粒子と、磁性体とを結合することで生成される結合物である。糖鎖は、アノマー炭素原子が置換を受けていない単糖鎖、オリゴ糖鎖、または、多糖鎖であり、還元末端を有する糖鎖には、濃縮対称のウイルスが結合する。リンカー化合物は、糖鎖と結合するアミノ基と、金属と結合する硫黄原子と、炭化水素鎖とを備えた化合物である。金属ナノ粒子は、金、銀、または、白金等の金属からなる微細な粒子であって、好ましくは１ｎｍ以上１００ｎｍ未満の平均粒子径を有するコロイド状の金属粒子である。磁性体は、例えば、酸化鉄、マグネタイト、または、フェライト等の磁性材料からなる。糖鎖固定化磁性粒子は、リガンド複合体と金属ナノ粒子と磁性体とを水等の溶媒中で混和することで生成できる。糖鎖固定化磁性粒子は、流動性を高めるために、水、生理食塩水、またはリン酸緩衝液等と混合して貯留室３３Ａに貯留されている。ウイルスを含む検体と、糖鎖固定化磁性粒子とを混合することで、ウイルスが糖鎖固定化磁性粒子により捕捉される。   The sugar chain-immobilized magnetic particles stored in the storage chamber 33A are metal particles having magnetism and having sugar chains immobilized thereon. Specifically, the sugar chain-immobilized magnetic particles are a conjugate produced by binding a ligand complex composed of a sugar chain and a linker compound, metal nanoparticles, and a magnetic substance. The sugar chain is a monosaccharide chain, an oligosaccharide chain, or a polysaccharide chain in which the anomeric carbon atom is not substituted, and a concentrated symmetrical virus binds to the sugar chain having a reducing end. The linker compound is a compound having an amino group that binds to a sugar chain, a sulfur atom that bonds to a metal, and a hydrocarbon chain. The metal nanoparticles are fine particles made of a metal such as gold, silver, or platinum, and are preferably colloidal metal particles having an average particle diameter of 1 nm or more and less than 100 nm. The magnetic body is made of a magnetic material such as iron oxide, magnetite, or ferrite, for example. The sugar chain-immobilized magnetic particles can be produced by mixing the ligand complex, metal nanoparticles, and magnetic material in a solvent such as water. The sugar chain-immobilized magnetic particles are mixed with water, physiological saline, phosphate buffer, or the like and stored in the storage chamber 33A in order to improve fluidity. By mixing the specimen containing the virus and the sugar chain-immobilized magnetic particles, the virus is captured by the sugar chain-immobilized magnetic particles.

貯留室３３Ｂが貯留する殺ウイルス剤は、ウイルスを破壊して不活化する薬剤である。具体的には、殺ウイルス剤は、例えば、ドデシル硫酸ナトリウム（ＳＤＳ）等の界面活性剤である。殺ウイルス剤は、流動性を高めるために、水等の溶媒と混合して貯留室３３Ａに貯留することができる。ウイルスを捕捉した糖鎖固定化磁性粒子と、殺ウイルス剤とを混合することにより、ウイルスの核酸が精製される。   The virucidal agent stored in the storage chamber 33B is a drug that destroys the virus and inactivates it. Specifically, the virucidal agent is a surfactant such as sodium dodecyl sulfate (SDS). The virusicide can be mixed with a solvent such as water and stored in the storage chamber 33A in order to improve fluidity. The nucleic acid of the virus is purified by mixing the sugar chain-immobilized magnetic particles that have captured the virus with a virucidal agent.

貯留室３３Ｃが貯留する希釈用の水は、超純水または蒸留水等の純度の高い水である。精製された核酸と、希釈用の水とを混合することにより、核酸を含む溶媒が所望の量となり、核酸の濃度が調整される。   The dilution water stored in the storage chamber 33C is high-purity water such as ultrapure water or distilled water. By mixing the purified nucleic acid and water for dilution, the solvent containing the nucleic acid becomes a desired amount, and the concentration of the nucleic acid is adjusted.

混合室１１が貯留する磁性粒子は、糖鎖固定化磁性粒子よりも平均粒子径の大きい磁性体である。磁性粒子は、糖鎖固定化磁性粒子を構成する磁性体と同様に、例えば、酸化鉄、マグネタイト、または、フェライト等の磁性材料からなる。   The magnetic particles stored in the mixing chamber 11 are magnetic bodies having a larger average particle diameter than the sugar chain-immobilized magnetic particles. The magnetic particles are made of a magnetic material such as iron oxide, magnetite, or ferrite, for example, in the same manner as the magnetic body constituting the sugar chain-immobilized magnetic particles.

また、貯留室３３Ａ〜３３Ｃには空気が含まれているものとする。蓋３０Ｂで密封されている貯留室３３Ａ〜３３Ｃ内の空気が膨張することによって、貯留室３３Ａ〜３３Ｃ内の物質を、混合室１１に吸入することができる。なお、貯留室３３Ａ〜３３Ｃの気密性が高くない場合には、回転部材３０の外部から貯留室３３Ａ〜３３Ｃに空気が流入することで、貯留室３３Ａ〜３３Ｃ内の物質が混合室１１に吸入される。   In addition, it is assumed that the storage chambers 33A to 33C contain air. When the air in the storage chambers 33A to 33C sealed by the lid 30B expands, the substance in the storage chambers 33A to 33C can be sucked into the mixing chamber 11. When the airtightness of the storage chambers 33A to 33C is not high, the air in the storage chambers 33A to 33C flows into the storage chambers 33A to 33C from the outside of the rotating member 30, so that the substances in the storage chambers 33A to 33C are sucked into the mixing chamber 11. Is done.

また、本明細書における「平均粒子径」は、複数の粒子に係る粒子径の平均値であって、「粒子径」は、粒子の二次元形状に対する最大内接円の直径とする。具体的には、例えば、粒子の二次元形状が楕円形状である場合は、その楕円の短径を粒子径とする。   The “average particle diameter” in the present specification is an average value of particle diameters related to a plurality of particles, and “particle diameter” is the diameter of the maximum inscribed circle with respect to the two-dimensional shape of the particles. Specifically, for example, when the two-dimensional shape of the particle is an ellipse, the minor axis of the ellipse is used as the particle diameter.

本実施形態の核酸精製方法では、下記の工程Ｓ１〜Ｓ１５を順番に経ることで、検体に含まれるウイルスの核酸を精製する。
・工程Ｓ１：検体を第１貯留室に入れる工程
・工程Ｓ２：第１貯留室と混合室とを連通状態にする工程
・工程Ｓ３：糖鎖固定化磁性粒子を混合室に吸入する工程
・工程Ｓ４：貯留室と混合室とを非連通状態にする工程
・工程Ｓ５：磁石部材を往復移動させる工程
・工程Ｓ６：第１貯留室と混合室とを連通状態にする工程
・工程Ｓ７：上清を第１貯留室に吐出する工程
・工程Ｓ８：第２貯留室と混合室とを連通状態にする工程
・工程Ｓ９：殺ウイルス剤を混合室に吸入する工程
・工程Ｓ１０：第３貯留室と混合室とを連通状態にする工程
・工程Ｓ１１：水を混合室に吸入する工程
・工程Ｓ１２：貯留室と混合室とを非連通状態にする工程
・工程Ｓ１３：膜部材に排出孔を形成する工程
・工程Ｓ１４：排出孔にノズルを取り付ける工程
・工程Ｓ１５：混合室から核酸を排出する工程 In the nucleic acid purification method of the present embodiment, viral nucleic acid contained in a sample is purified by sequentially performing the following steps S1 to S15.
-Step S1: Step of putting the specimen into the first storage chamber-Step S2: Step of bringing the first storage chamber and the mixing chamber into communication with each other-Step S3: Step of sucking the sugar chain-immobilized magnetic particles into the mixing chamber S4: Steps for bringing the storage chamber and the mixing chamber into a non-communication state. Step S5: Steps for reciprocating the magnet member. Step S6: Steps for bringing the first storage chamber and the mixing chamber into communication. Step S7: Supernatant Step S8: Step of making the second storage chamber and the mixing chamber communicate with each other Step S9: Step of sucking the virusicide into the mixing chamber Step S10: Step S11 for bringing the mixing chamber into communication state Step S11: Step for sucking water into the mixing chamber Step S12 Step for making communication between the storage chamber and the mixing chamber Step S13: Forming a discharge hole in the membrane member Step / step S14: Step / step S1 for attaching the nozzle to the discharge hole : Step of discharging the nucleic acid from the mixing chamber

工程Ｓ１では、カバー部材７０および蓋３０Ｂを開き、糖鎖固定化磁性粒子を貯留する貯留室３３Ａに、例えばマイクロピペットを用いてウイルスを含んだ唾液を入れる。こうして、工程Ｓ１では、唾液と糖鎖固定化磁性粒子とが混合され、糖鎖固定化磁性粒子が、唾液に含まれるウイルスを捕捉する。次いで、工程Ｓ２では、貯留室３３Ａと混合室１１とが連通状態となるように回転部材３０を回転させる。こうして、工程Ｓ２では、貯留室３３Ａと混合室１１とを連通状態にして、貯留室３３Ａ内の物質を混合室１１に吸入可能な状態となる。   In step S1, the cover member 70 and the lid 30B are opened, and the saliva containing the virus is put into the storage chamber 33A storing the sugar chain-immobilized magnetic particles using, for example, a micropipette. Thus, in step S1, saliva and sugar chain-immobilized magnetic particles are mixed, and the sugar chain-immobilized magnetic particles capture the virus contained in the saliva. Next, in step S2, the rotating member 30 is rotated so that the storage chamber 33A and the mixing chamber 11 are in communication with each other. Thus, in step S2, the storage chamber 33A and the mixing chamber 11 are brought into communication with each other, and the substance in the storage chamber 33A can be sucked into the mixing chamber 11.

次いで、工程Ｓ３では、混合室１１内の容積が拡大するようにピストン部材４０を摺動させる（引き上げる）ことで、ウイルスを捕捉した糖鎖固定化磁性粒子を、貯留室３３Ａから混合室１１に吸入する。こうして、工程Ｓ３では、ウイルスを捕捉している糖鎖固定化磁性粒子と、当該磁性粒子よりも平均粒子径の大きい磁性粒子とが混合され、糖鎖固定化磁性粒子が磁力で吸引され易くなる。なお、磁性粒子が十分に混合されるように、磁石部材５０を混合室１１の上端部近傍に配置しておくこと、または、ベース部材１０から磁石部材５０を取り外しておくことが好ましい。   Next, in step S3, the piston member 40 is slid (pulled up) so that the volume in the mixing chamber 11 is expanded, whereby the sugar chain-immobilized magnetic particles that have captured the virus are transferred from the storage chamber 33A to the mixing chamber 11. Inhale. Thus, in step S3, the sugar chain-immobilized magnetic particles capturing the virus and the magnetic particles having a larger average particle diameter than the magnetic particles are mixed, and the sugar chain-immobilized magnetic particles are easily attracted by magnetic force. . In addition, it is preferable to arrange the magnet member 50 in the vicinity of the upper end portion of the mixing chamber 11 or to remove the magnet member 50 from the base member 10 so that the magnetic particles are sufficiently mixed.

次いで、工程Ｓ４では、貯留室３３Ａ〜３３Ｃと混合室１１とが非連通状態になるように回転部材３０を回転させる。こうして、工程Ｓ４では、貯留室３３Ａ〜３３Ｃと混合室１１とを非連通状態にして、混合室１１内に物質が密閉された状態となる。次いで、工程Ｓ５では、混合室１１内で検体と磁性粒子とを十分に混合するために、ベース部材１０に対して磁石部材５０を上下方向に往復移動させる。こうして、工程Ｓ５では、磁石部材５０の移動によって磁性粒子と検体とを十分に混合し、ウイルスを捕捉している糖鎖固定化磁性粒子を混合室１１内で凝集させる。   Next, in step S4, the rotation member 30 is rotated so that the storage chambers 33A to 33C and the mixing chamber 11 are not in communication. Thus, in step S4, the storage chambers 33A to 33C and the mixing chamber 11 are brought into a non-communication state, and the substance is sealed in the mixing chamber 11. Next, in step S <b> 5, the magnet member 50 is reciprocated in the vertical direction with respect to the base member 10 in order to sufficiently mix the specimen and the magnetic particles in the mixing chamber 11. Thus, in step S5, the magnetic particles and the specimen are sufficiently mixed by the movement of the magnet member 50, and the sugar chain-immobilized magnetic particles capturing the virus are aggregated in the mixing chamber 11.

次いで、工程Ｓ６では、工程Ｓ２と同様に、貯留室３３Ａと混合室１１とが連通状態となるように回転部材３０を回転させる。こうして、工程Ｓ６では、貯留室３３Ａと混合室１１とを連通状態にして、混合室１１内の物質を貯留室３３Ａに吐出可能な状態となる。次いで、工程Ｓ７では、磁石部材５０により混合室１１内に磁場を発生させた状態で、混合室１１内の容積が縮小するようにピストン部材４０を摺動させる（押し下げる）ことで、混合室１１内の上清を貯留室３３Ａに吐出する。このとき、糖鎖固定化磁性粒子が混合室１１から流出しないように、磁石部材５０は、貫通孔１３が設けられた混合室１１の下端部近傍に配置しておくことが好ましい。こうして、工程Ｓ７では、混合室１１内の上清が貯留室３３Ａに吐出され、ウイルスを捕捉している磁性粒子は貯留室３３Ａに吐出されずに混合室１１に留まり、混合室１１内でウイルスが濃縮された状態となる。   Next, in step S6, as in step S2, the rotating member 30 is rotated so that the storage chamber 33A and the mixing chamber 11 are in communication with each other. Thus, in step S6, the storage chamber 33A and the mixing chamber 11 are brought into communication with each other, so that the substance in the mixing chamber 11 can be discharged into the storage chamber 33A. Next, in step S7, in a state where a magnetic field is generated in the mixing chamber 11 by the magnet member 50, the piston member 40 is slid (pressed down) so that the volume in the mixing chamber 11 is reduced, thereby mixing the chamber 11. The inner supernatant is discharged into the storage chamber 33A. At this time, the magnet member 50 is preferably arranged in the vicinity of the lower end of the mixing chamber 11 in which the through-hole 13 is provided so that the sugar chain-immobilized magnetic particles do not flow out of the mixing chamber 11. Thus, in step S7, the supernatant in the mixing chamber 11 is discharged into the storage chamber 33A, and the magnetic particles capturing the virus remain in the mixing chamber 11 without being discharged into the storage chamber 33A. Becomes a concentrated state.

次いで、工程Ｓ８では、貯留室３３Ｂと混合室１１とが連通状態となるように回転部材３０を回転させる。こうして、工程Ｓ８では、貯留室３３Ｂと混合室１１とを連通状態にして、貯留室３３Ｂ内の物質を混合室１１に吸入可能な状態となる。次いで、工程Ｓ９では、混合室１１内の容積が拡大するようにピストン部材４０を摺動させることで、殺ウイルス剤を、貯留室３３Ｂから混合室１１に吸入する。なお、糖鎖固定化磁性粒子と殺ウイルス剤が十分に混合されるように、磁石部材５０を混合室１１の上端部近傍に配置しておくこと、または、ベース部材１０から磁石部材５０を取り外しておくことが好ましい。こうして、工程Ｓ９では、殺ウイルス剤と、ウイルスを捕捉している糖鎖固定化磁性粒子とが混合され、ウイルスが破壊され、糖鎖固定化磁性粒子からウイルスの核酸が分離した状態となる。   Next, in step S8, the rotating member 30 is rotated so that the storage chamber 33B and the mixing chamber 11 are in communication with each other. Thus, in step S8, the storage chamber 33B and the mixing chamber 11 are brought into communication with each other, and the substance in the storage chamber 33B can be sucked into the mixing chamber 11. Next, in step S9, the virucidal agent is sucked into the mixing chamber 11 from the storage chamber 33B by sliding the piston member 40 so that the volume in the mixing chamber 11 increases. In addition, the magnet member 50 is disposed in the vicinity of the upper end of the mixing chamber 11 so that the sugar chain-immobilized magnetic particles and the virusicide are sufficiently mixed, or the magnet member 50 is removed from the base member 10. It is preferable to keep it. Thus, in step S9, the virucidal agent and the sugar chain-immobilized magnetic particles capturing the virus are mixed, the virus is destroyed, and the virus nucleic acid is separated from the sugar chain-immobilized magnetic particles.

次いで、工程Ｓ１０では、貯留室３３Ｃと混合室１１とが連通状態となるように回転部材３０を回転させる。こうして、工程Ｓ１０では、貯留室３３Ｃと混合室１１とを連通状態にして、貯留室３３Ｃ内の物質を混合室１１に吸入可能な状態となる。次いで、工程Ｓ１１では、混合室１１内の容積が拡大するようにピストン部材４０を摺動させることで、希釈用の水を、貯留室３３Ｃから混合室１１に吸入する。こうして、工程Ｓ１１では、水と、磁性粒子およびウイルスの核酸とが混合され、ウイルスの核酸の濃度が調整される。次いで、工程Ｓ１２では、工程Ｓ４と同様に、貯留室３３Ａ〜３３Ｃと混合室１１とが非連通状態になるように回転部材３０を回転させる。こうして、工程Ｓ１２では、貯留室３３Ａ〜３３Ｃと混合室１１とを非連通状態にして、混合室１１内の物質が貯留室３３Ａ〜３３Ｃに吐出されない状態となる。   Next, in step S10, the rotating member 30 is rotated so that the storage chamber 33C and the mixing chamber 11 are in communication with each other. Thus, in step S10, the storage chamber 33C and the mixing chamber 11 are brought into communication with each other, and the substance in the storage chamber 33C can be sucked into the mixing chamber 11. Next, in step S11, the piston member 40 is slid so that the volume in the mixing chamber 11 is increased, whereby dilution water is sucked into the mixing chamber 11 from the storage chamber 33C. Thus, in step S11, water, magnetic particles, and viral nucleic acid are mixed, and the concentration of viral nucleic acid is adjusted. Next, in step S12, as in step S4, the rotating member 30 is rotated so that the storage chambers 33A to 33C and the mixing chamber 11 are not in communication. Thus, in step S12, the storage chambers 33A to 33C and the mixing chamber 11 are disconnected from each other, and the substance in the mixing chamber 11 is not discharged into the storage chambers 33A to 33C.

次いで、工程Ｓ１３では、排出用ノズル９０の先端部９２で弾性部材２０の膜部２２を穿つことによって、弾性部材２０に排出孔（図示略）を形成し、工程Ｓ１４では、排出用ノズル９０の基端部９１を弾性部材２０のノズル保持部２４に圧入することによって、工程Ｓ１３で形成した排出孔に排出用ノズル９０を取り付ける。こうして、工程Ｓ１３およびＳ１４では、混合室１１内の物質を器具本体１Ａの外部に排出可能な状態とする。   Next, in step S13, a discharge hole (not shown) is formed in the elastic member 20 by piercing the film portion 22 of the elastic member 20 with the tip 92 of the discharge nozzle 90. In step S14, the discharge nozzle 90 By pressing the base end portion 91 into the nozzle holding portion 24 of the elastic member 20, the discharge nozzle 90 is attached to the discharge hole formed in step S13. Thus, in steps S13 and S14, the substance in the mixing chamber 11 is brought into a state where it can be discharged to the outside of the instrument body 1A.

次いで、工程Ｓ１５では、磁石部材５０により混合室１１内に磁場を発生させた状態で、混合室１１内の容積が縮小するようにピストン部材４０を摺動させることで、ウイルスの核酸を含んだ溶液（懸濁液）を混合室１１から所定量だけ排出する。なお、混合室１１内の溶液を所定量だけ精確に採取するために、排出用ノズル９０内の全ての空気を先端部９２から事前に排出しておくことが好ましい。また、工程Ｓ１５では、摺動制限部材６０を用いてピストン部材４０の摺動を制限することで、混合室１１内の溶液を所定量だけ精確に排出する。また、磁性粒子が混合室１１から流出しないように、磁石部材５０は、排出孔が設けられた混合室１１の下端部近傍に配置しておくことが好ましい。こうして、工程Ｓ１５では、ウイルスの核酸を含んだ上清が、混合室１１から排出用ノズル９０を通して排出される。   Next, in step S15, in the state in which a magnetic field is generated in the mixing chamber 11 by the magnet member 50, the piston member 40 is slid so that the volume in the mixing chamber 11 is reduced. A predetermined amount of the solution (suspension) is discharged from the mixing chamber 11. In order to accurately collect a predetermined amount of the solution in the mixing chamber 11, it is preferable to discharge all the air in the discharge nozzle 90 from the tip portion 92 in advance. In step S15, the sliding limit member 60 is used to limit the sliding of the piston member 40, whereby the solution in the mixing chamber 11 is accurately discharged by a predetermined amount. Moreover, it is preferable to arrange the magnet member 50 in the vicinity of the lower end of the mixing chamber 11 provided with the discharge hole so that the magnetic particles do not flow out of the mixing chamber 11. Thus, in step S15, the supernatant containing the viral nucleic acid is discharged from the mixing chamber 11 through the discharge nozzle 90.

以上のようにして精製されたウイルスの核酸をＰＣＲ法で解析することによって、唾液に含まれていたウイルスを同定および定量することができる。具体的には、例えば、精製された核酸がＲＮＡであれば、リアルタイム逆転写ポリメラーゼ連鎖反応によりウイルスを同定および定量することができる。   By analyzing the nucleic acid of the virus purified as described above by the PCR method, the virus contained in the saliva can be identified and quantified. Specifically, for example, if the purified nucleic acid is RNA, the virus can be identified and quantified by real-time reverse transcription polymerase chain reaction.

本実施形態においては以下の効果が得られる。
（１）回転部材３０の回転角が変化することで、貯留室３３Ａ〜３３Ｃと混合室１１との連通状態が切り替わる。また、ピストン部材４０が摺動することで、混合室１１の内部の容積が変化する。このため、貯留室３３Ａ〜３３Ｃの各々から混合室１１に物質を順番に吸入することで、複数の物質を混合することができる。したがって、核酸精製キット１の使用者は、貯留室３３Ａに唾液を入れた後、ベース部材１０に対して回転部材３０を回転させる操作と、ベース部材１０に対してピストン部材４０を摺動させる操作を行うことで、糖鎖固定化磁性粒子や殺ウイルス剤等の複数の物質を簡単に混合することができる。その結果、ウイルスの核酸を容易に精製することができる。 In the present embodiment, the following effects can be obtained.
(1) The communication state between the storage chambers 33 </ b> A to 33 </ b> C and the mixing chamber 11 is switched by changing the rotation angle of the rotating member 30. Moreover, the volume of the inside of the mixing chamber 11 changes because the piston member 40 slides. For this reason, a plurality of substances can be mixed by sequentially sucking substances into the mixing chamber 11 from each of the storage chambers 33A to 33C. Therefore, the user of the nucleic acid purification kit 1 puts saliva into the storage chamber 33 </ b> A, then rotates the rotating member 30 with respect to the base member 10, and slides the piston member 40 with respect to the base member 10. By performing the above, it is possible to easily mix a plurality of substances such as sugar chain-immobilized magnetic particles and virucidal agents. As a result, viral nucleic acid can be easily purified.

（２）磁石部材５０が、ベース部材１０に対して摺動可能に設けられているため、磁石部材５０が動かない構成に比べて、混合室１１内で検体と磁性粒子とを十分に混合することができる。その結果、ウイルスを捕捉した糖鎖固定化磁性粒子を効率良く凝集させ、ウイルスを効率良く濃縮できる。   (2) Since the magnet member 50 is provided so as to be slidable with respect to the base member 10, the sample and the magnetic particles are sufficiently mixed in the mixing chamber 11 as compared with the configuration in which the magnet member 50 does not move. be able to. As a result, the sugar chain-immobilized magnetic particles capturing the virus can be efficiently aggregated and the virus can be efficiently concentrated.

（３）摺動制限部材６０は、ピストンロッド４２に設けられた突部４４に当接することで、ピストン部材４０の摺動を制限するため、混合室１１内の容積の変化を制限することができる。その結果、混合室１１内の物質を所望量だけ混合室１１から排出することができる。   (3) Since the sliding restriction member 60 is in contact with the protrusion 44 provided on the piston rod 42 to restrict the sliding of the piston member 40, the change in the volume in the mixing chamber 11 can be restricted. it can. As a result, the substance in the mixing chamber 11 can be discharged from the mixing chamber 11 by a desired amount.

（４）排出用ノズル９０で弾性部材２０（膜部材）が穿たれることにより排出孔が形成されるため、専用の道具を用いることなく弾性部材２０に排出孔を形成することができる。また、排出孔が形成されるまでは混合室１１の密閉性を確保することができる。   (4) Since the discharge hole is formed by the elastic member 20 (membrane member) being pierced by the discharge nozzle 90, the discharge hole can be formed in the elastic member 20 without using a dedicated tool. Moreover, the sealing property of the mixing chamber 11 can be ensured until the discharge hole is formed.

（５）貯留室３３Ａ〜３３Ｃおよび混合室１１が密閉された状態となるため、物質を混合する際に異物が混入することを防ぐことができる。このため、コンタミネーションを防ぎ、クリーンルーム外において核酸を容易に精製することができる。   (5) Since the storage chambers 33 </ b> A to 33 </ b> C and the mixing chamber 11 are sealed, it is possible to prevent foreign matters from being mixed when substances are mixed. For this reason, contamination can be prevented and the nucleic acid can be easily purified outside the clean room.

（６）上記の核酸精製キット１を用いた核酸精製方法によれば、ウイルスを捕捉している糖鎖固定化磁性粒子と、当該磁性粒子よりも平均粒子径の大きい磁性粒子とを混合することで、ウイルスの濃縮効率を向上させることができる。このため、遠心分離機等の大掛かりな装置を必要とせずに、検体に含まれる微量のウイルスから核酸を精製することができる。   (6) According to the nucleic acid purification method using the nucleic acid purification kit 1, the sugar chain-immobilized magnetic particles capturing the virus are mixed with the magnetic particles having an average particle size larger than that of the magnetic particles. Thus, the virus concentration efficiency can be improved. For this reason, a nucleic acid can be purified from a very small amount of virus contained in a specimen without requiring a large-scale apparatus such as a centrifuge.

本発明は、上記実施形態に限定されるものではなく、上記構成を変更することもできる。例えば、以下のように変更して実施することもでき、以下の変更を組み合わせて実施することもできる。   The present invention is not limited to the above embodiment, and the above configuration can be changed. For example, the following modifications can be implemented, and the following modifications can be combined.

・上記実施形態では弾性部材２０が軸受部材および膜部材を兼ねているが、軸受部材および膜部材を、それぞれエラストマーにより形成された別部品により構成することもできる。   In the above embodiment, the elastic member 20 serves as both a bearing member and a film member. However, the bearing member and the film member may be configured by separate parts each formed of an elastomer.

・上記実施形態では斜めに形成された突部４４および直線状の突部４５がピストンロッド４２に設けられているが、例えば図１３に示すように、ピストンロッドに設けられる突部の形状を適宜変更してもよい。また、例えば図１４に示すように、ピストンロッドの突部の形状に合わせて、摺動制限部材の形状を適宜変更してもよい。   In the above embodiment, the obliquely formed protrusion 44 and the linear protrusion 45 are provided on the piston rod 42. For example, as shown in FIG. 13, the shape of the protrusion provided on the piston rod is appropriately set. It may be changed. Further, for example, as shown in FIG. 14, the shape of the sliding restriction member may be appropriately changed in accordance with the shape of the protrusion of the piston rod.

図１３を参照して、変形例に係るピストン部材１４０について説明する。なお、上記実施形態に係るピストン部材４０と同様の構成については説明を省略する。
ピストン部材１４０は、ピストンヘッド４１と、ピストンロッド４２と、リング４３と、ピストンロッド４２の側部に設けられた突部４６とを有している。突部４６は、ピストンロッド４２が延びる上下方向において所定間隔をあけて複数設けられ、円柱状に形成されている。 With reference to FIG. 13, the piston member 140 which concerns on a modification is demonstrated. In addition, description is abbreviate | omitted about the structure similar to the piston member 40 which concerns on the said embodiment.
The piston member 140 includes a piston head 41, a piston rod 42, a ring 43, and a protrusion 46 provided on a side portion of the piston rod 42. A plurality of protrusions 46 are provided at predetermined intervals in the vertical direction in which the piston rod 42 extends, and are formed in a columnar shape.

図１４を参照して、変形例に係る摺動制限部材１６０について説明する。なお、上記実施形態に係る摺動制限部材６０と同様の構成については説明を省略する。また、図１４（Ｂ）は、図１４（Ａ）のＥ−Ｅ線断面図である。
摺動制限部材１６０は、基部６１と、左右一対の突出部６２と、ベース部材係合部６４と、左右一対の突出部６２の内側部分に設けられたピストン部材係合部６５とを有している。ピストン部材係合部６５は、ピストン部材１４０の複数の突部４６の間に挿し込まれ、突部４６と当接することによって、ピストン部材１４０の上下方向の移動を規制する。ピストン部材係合部６５は、所定の厚みを有する第１段部６５Ａと、第１段部６５Ａよりも厚みが小さい第２段部６５Ｂとを有している。 With reference to FIG. 14, the sliding restriction member 160 which concerns on a modification is demonstrated. In addition, description is abbreviate | omitted about the structure similar to the sliding limiting member 60 which concerns on the said embodiment. FIG. 14B is a cross-sectional view taken along line EE in FIG.
The sliding restriction member 160 includes a base 61, a pair of left and right protrusions 62, a base member engagement part 64, and a piston member engagement part 65 provided on an inner portion of the pair of left and right protrusions 62. ing. The piston member engaging portion 65 is inserted between the plurality of protrusions 46 of the piston member 140 and abuts against the protrusions 46 to restrict the movement of the piston member 140 in the vertical direction. The piston member engaging portion 65 has a first step portion 65A having a predetermined thickness and a second step portion 65B having a thickness smaller than that of the first step portion 65A.

ピストン部材１４０は、複数の突部４６の間に摺動制限部材１６０のピストン部材係合部６５が挿し込まれることによって、上下方向における摺動が規制される。ピストン部材１４０を上下方向に摺動させるためには、複数の突部４６の間からピストン部材係合部６５が抜け出るように摺動制限部材１６０を後方に移動させて、ピストン部材１４０の摺動の制限を解除する。この変形例においては、上記の工程Ｓ１５で、混合室１１内の溶液を所定量だけ精確に排出するために、突部４６を第１段部６５Ａに当接させ、ピストン部材１４０を下方に押さえながら摺動制限部材１６０を前方に移動させることで、突部４６を第２段部６５Ｂに当接させる。このようにして、第１段部６５Ａと第２段部６５Ｂとの段差分だけ、ピストン部材１４０が下方に摺動して、排出孔から核酸を含む所定量の溶液が排出される。   The piston member 140 is restricted from sliding in the vertical direction by inserting the piston member engaging portion 65 of the sliding restriction member 160 between the plurality of protrusions 46. In order to slide the piston member 140 in the vertical direction, the sliding limit member 160 is moved backward so that the piston member engaging portion 65 comes out from between the plurality of protrusions 46, and the piston member 140 slides. Remove the restrictions. In this modified example, in order to accurately discharge a predetermined amount of the solution in the mixing chamber 11 by the above-described step S15, the protrusion 46 is brought into contact with the first step portion 65A and the piston member 140 is pressed downward. However, the protrusion 46 is brought into contact with the second step portion 65B by moving the sliding restriction member 160 forward. In this manner, the piston member 140 slides downward by an amount corresponding to the step between the first step portion 65A and the second step portion 65B, and a predetermined amount of solution containing nucleic acid is discharged from the discharge hole.

・上記実施形態では、ベース部材保持部５３がベース部材１０の後端部を挟むことでベース部材１０に磁石部材５０が取り付けられていたが、ベース部材１０に対する磁石部材５０の取付構造を適宜変更してもよい。例えば、ベース部材の後端部に上下方向に延びる凹状の溝が形成され、当該溝に磁石部材の一部が圧入されることで、ベース部材に磁石部材が取り付けられてもよい。   In the above embodiment, the magnet member 50 is attached to the base member 10 with the base member holding portion 53 sandwiching the rear end portion of the base member 10, but the attachment structure of the magnet member 50 to the base member 10 is appropriately changed. May be. For example, a concave groove extending in the vertical direction may be formed in the rear end portion of the base member, and the magnet member may be attached to the base member by pressing a part of the magnet member into the groove.

・上記実施形態では磁石部材５０がベース部材１０に対して摺動可能に設けられているが、混合室１１内で検体と磁性粒子とを十分に混合することができるのであれば、磁石部材５０は、ベース部材１０に対して摺動不可能とすることもできる。   In the above embodiment, the magnet member 50 is provided so as to be slidable with respect to the base member 10. However, if the specimen and the magnetic particles can be sufficiently mixed in the mixing chamber 11, the magnet member 50. Can be made non-slidable with respect to the base member 10.

・上記実施形態では回転部材３０が３つの貯留室３３Ａ〜３３Ｃを有していたが、回転部材が２つまたは４つ以上の貯留室を有していてもよい。具体的には、例えば、希釈用の水を貯留する貯留室３３Ｃを省略してもよい。この場合、上記工程Ｓ１０，Ｓ１１を省略した核酸精製方法によって、混合室内の核酸の濃度調整を行わずに核酸を精製することができる。また、例えば、検体として糞尿を用いる場合に備えて、検体を洗浄するための水またはリン酸緩衝液等を貯留する第４貯留室を回転部材が有していてもよい。すなわち、回転部材は複数の貯留室を有していればよい。   In the above embodiment, the rotating member 30 has the three storage chambers 33A to 33C, but the rotating member may have two or four or more storage chambers. Specifically, for example, the storage chamber 33 </ b> C for storing dilution water may be omitted. In this case, the nucleic acid can be purified without adjusting the concentration of the nucleic acid in the mixing chamber by the nucleic acid purification method in which the steps S10 and S11 are omitted. In addition, for example, the rotating member may have a fourth storage chamber for storing water or a phosphate buffer solution for washing the specimen in preparation for using manure as the specimen. That is, the rotating member only needs to have a plurality of storage chambers.

・上記実施形態では、ベース部材１０に１つの目印１５が形成されるとともに、回転部材３０の基板部３１に３つの目印３７Ａ〜３７Ｃが形成されているが、貯留室と混合室との連通状態を確認することが可能な目印の個数、配置箇所、形状を適宜変更してもよい。   In the above embodiment, one mark 15 is formed on the base member 10 and three marks 37A to 37C are formed on the substrate portion 31 of the rotating member 30, but the communication state between the storage chamber and the mixing chamber The number, location, and shape of the marks that can be confirmed may be appropriately changed.

・核酸精製キット１を構成する部品の形状や材料を適宜変更してもよい。また、上記実施形態ではウイルスを含む検体として唾液を用いるものであるが、鼻粘膜、動物の体液、糞尿等を検体として用いてもよい。   -You may change suitably the shape and material of the components which comprise the nucleic acid purification kit 1. FIG. In the above-described embodiment, saliva is used as a specimen containing a virus, but nasal mucosa, animal body fluid, feces and urine may be used as a specimen.

・混合室１１の磁性粒子の流動性を高めるために、磁性粒子を、水、生理食塩水、またはリン酸緩衝液等と混合した状態で、混合室１１に貯留してもよい。また、検体を入れることで糖鎖固定化磁性粒子の流動性を確保することができるのであれば、糖鎖固定化磁性粒子は、水、生理食塩水、またはリン酸緩衝液等と混合せずに貯留室３３Ａに貯留してもよい。   In order to increase the fluidity of the magnetic particles in the mixing chamber 11, the magnetic particles may be stored in the mixing chamber 11 in a state of being mixed with water, physiological saline, phosphate buffer, or the like. In addition, if the fluidity of the sugar chain-immobilized magnetic particles can be ensured by adding a sample, the sugar chain-immobilized magnetic particles are not mixed with water, physiological saline, phosphate buffer, or the like. Alternatively, it may be stored in the storage chamber 33A.

・上記実施形態では本発明の混合器具は核酸精製キット１であるが、本発明は、核酸を精製する器具に限られず、任意の複数の物質を順に混合する器具として適宜使用することができる。   -In the said embodiment, although the mixing instrument of this invention is the nucleic acid purification kit 1, this invention is not restricted to the instrument which refine | purifies a nucleic acid, It can use suitably as an instrument which mixes arbitrary several substances in order.

１ 核酸精製キット（混合器具）
１Ａ 器具本体
１０ ベース部材
１１ 混合室
２０ 弾性部材（膜部材）
３０ 回転部材
３３Ａ 貯留室（第１貯留室）
３３Ｂ 貯留室（第２貯留室）
３３Ｃ 貯留室（第３貯留室）
４０，１４０ ピストン部材
４１ ピストンヘッド
４２ ピストンロッド
４４，４５，４６ 突部
５０ 磁石部材
６０，１６０ 摺動制限部材
７０ カバー部材
９０ 排出用ノズル 1 Nucleic acid purification kit (mixing device)
1A Instrument body 10 Base member 11 Mixing chamber 20 Elastic member (membrane member)
30 rotating member 33A storage chamber (first storage chamber)
33B Reservoir (second reservoir)
33C Reservoir (third reservoir)
40,140 Piston member 41 Piston head 42 Piston rods 44, 45, 46 Protrusion 50 Magnet member 60, 160 Sliding limiting member 70 Cover member 90 Discharging nozzle

Claims (5)

複数の物質を混合する混合器具であって、
混合室を有したベース部材と、
前記ベース部材に対して回転可能に設けられた回転部材と、
前記ベース部材に対して摺動可能に設けられたピストン部材とを備え、
前記回転部材は、前記物質を貯留するための複数の貯留室を有し、
前記貯留室は、前記回転部材の回転角が特定の角度であるとき前記混合室と連通するように構成されており、前記回転部材の回転角を変化させることで、前記貯留室と前記混合室との連通状態が切り替わり、
前記ピストン部材は、前記混合室を閉塞するピストンヘッドを有しており、前記ピストン部材を摺動させることで、前記混合室の内部の容積が変化する
ことを特徴とする混合器具。 A mixing device for mixing a plurality of substances,
A base member having a mixing chamber;
A rotating member provided rotatably with respect to the base member;
A piston member slidably provided with respect to the base member,
The rotating member has a plurality of storage chambers for storing the substance,
The storage chamber is configured to communicate with the mixing chamber when the rotation angle of the rotating member is a specific angle, and the storage chamber and the mixing chamber are changed by changing the rotation angle of the rotating member. The communication status with
The said piston member has a piston head which obstruct | occludes the said mixing chamber, The volume inside the said mixing chamber changes by sliding the said piston member. The mixing instrument characterized by the above-mentioned. 前記混合室の内部に磁場を発生させる磁石部材をさらに備え、
前記磁石部材は、前記ベース部材に対して摺動可能に設けられている
ことを特徴とする請求項１に記載の混合器具。 A magnetic member that generates a magnetic field inside the mixing chamber;
The mixing device according to claim 1, wherein the magnet member is slidable with respect to the base member. 前記ピストン部材の摺動を制限する摺動制限部材をさらに備え、
前記ピストン部材は、前記ベース部材の内部から外部に延びたピストンロッドを有し、当該ピストンロッドには、当該ピストンロッドが延びる方向において複数の突部が所定間隔をあけて設けられており、
前記摺動制限部材は、前記突部に当接することで、前記ピストン部材の摺動を制限する
ことを特徴とする請求項１または２に記載の混合器具。 A sliding limiting member that limits sliding of the piston member;
The piston member has a piston rod extending from the inside of the base member to the outside, and the piston rod is provided with a plurality of protrusions at predetermined intervals in a direction in which the piston rod extends,
The mixing device according to claim 1 or 2, wherein the sliding restriction member restricts sliding of the piston member by abutting against the protrusion. 前記混合室を閉塞する膜部材と、
前記膜部材を穿つことが可能な排出用ノズルとをさらに備え、
前記排出用ノズルは、前記膜部材が穿たれることで形成される排出孔に取り付けられる
ことを特徴とする請求項１〜３のいずれか一項に記載の混合器具。 A membrane member for closing the mixing chamber;
A discharge nozzle that can pierce the membrane member;
The mixing device according to any one of claims 1 to 3, wherein the discharge nozzle is attached to a discharge hole formed when the membrane member is pierced. 前記回転部材が少なくとも２つの前記貯留室を有し、ウイルスを捕捉する糖鎖固定化磁性粒子が前記貯留室としての第１貯留室に貯留され、ウイルスを不活化する殺ウイルス剤が前記貯留室としての第２貯留室に貯留され、前記糖鎖固定化磁性粒子よりも平均粒子径の大きい磁性粒子が前記混合室に貯留されている請求項１〜４のいずれか一項に記載の混合器具を用いた核酸精製方法であって、
ウイルスを含んだ検体を前記第１貯留室に入れる第１工程と、
前記第１工程の後に、前記第１貯留室と前記混合室とが連通状態となるように前記回転部材を回転させる第２工程と、
前記第２工程の後に、前記混合室の内部の容積が拡大するように前記ピストン部材を摺動させることで、前記検体に含まれる前記ウイルスを捕捉した前記糖鎖固定化磁性粒子を、前記第１貯留室から前記混合室に吸入する第３工程と、
前記第３工程の後に、前記混合室の内部に磁場を発生させた状態で、前記混合室の内部の容積が縮小するように前記ピストン部材を摺動させることで、前記混合室内の上清を前記第１貯留室に吐出する第４工程と、
前記第４工程の後に、前記第２貯留室と前記混合室とが連通状態となるように前記回転部材を回転させる第５工程と、
前記第５工程の後に、前記混合室の内部の容積が拡大するように前記ピストン部材を摺動させることで、前記殺ウイルス剤を、前記第２貯留室から前記混合室に吸入する第６工程と、
前記第６工程の後に、前記混合室の内部に磁場を発生させた状態で、前記混合室の内部の容積が縮小するように前記ピストン部材を摺動させることで、前記ウイルスの核酸を前記混合室から排出する第７工程とを含む
ことを特徴とする核酸精製方法。 The rotating member has at least two of the storage chambers, sugar chain-immobilized magnetic particles for capturing viruses are stored in the first storage chamber as the storage chamber, and a virusicide that inactivates the virus is the storage chamber. The mixing device according to any one of claims 1 to 4, wherein magnetic particles stored in the second storage chamber are stored in the mixing chamber and have a larger average particle diameter than the sugar chain-immobilized magnetic particles. A nucleic acid purification method using
A first step of placing a specimen containing a virus in the first reservoir;
After the first step, a second step of rotating the rotating member so that the first storage chamber and the mixing chamber are in communication with each other;
After the second step, the sugar chain-immobilized magnetic particles capturing the virus contained in the specimen are slid by sliding the piston member so that the internal volume of the mixing chamber is expanded. A third step of sucking from one reservoir into the mixing chamber;
After the third step, with the magnetic field generated inside the mixing chamber, the piston member is slid so that the volume inside the mixing chamber is reduced. A fourth step of discharging into the first storage chamber;
After the fourth step, a fifth step of rotating the rotating member so that the second storage chamber and the mixing chamber are in communication with each other;
After the fifth step, the sixth step of sucking the virusicide from the second storage chamber into the mixing chamber by sliding the piston member so that the volume inside the mixing chamber is enlarged. When,
After the sixth step, the nucleic acid of the virus is mixed by sliding the piston member so that the volume inside the mixing chamber is reduced in a state where a magnetic field is generated inside the mixing chamber. And a seventh step of discharging from the chamber.
JP2016082988A 2016-04-18 2016-04-18 Mixing device, and nucleic acid purification method Pending JP2017192310A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108261954A (en) * 2018-01-29 2018-07-10 重庆好弟兄食品有限公司 Food processing material blending device
WO2023071049A1 (en) * 2021-11-01 2023-05-04 江苏汇先医药技术有限公司 Vertical micro-fluidic chip and method for pcr test

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108261954A (en) * 2018-01-29 2018-07-10 重庆好弟兄食品有限公司 Food processing material blending device
WO2023071049A1 (en) * 2021-11-01 2023-05-04 江苏汇先医药技术有限公司 Vertical micro-fluidic chip and method for pcr test

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