JP2003500674A5 - - Google Patents
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- JP2003500674A5 JP2003500674A5 JP2000621073A JP2000621073A JP2003500674A5 JP 2003500674 A5 JP2003500674 A5 JP 2003500674A5 JP 2000621073 A JP2000621073 A JP 2000621073A JP 2000621073 A JP2000621073 A JP 2000621073A JP 2003500674 A5 JP2003500674 A5 JP 2003500674A5
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- cartridge
- reaction chamber
- chamber
- reaction
- flow path
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- 238000006243 chemical reaction Methods 0.000 description 43
- 239000012528 membrane Substances 0.000 description 7
- 239000012530 fluid Substances 0.000 description 6
- 230000009089 cytolysis Effects 0.000 description 5
- 230000002934 lysing Effects 0.000 description 5
- 230000003287 optical Effects 0.000 description 4
- 241000700605 Viruses Species 0.000 description 3
- 210000003284 Horns Anatomy 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000010408 film Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000010409 thin film Substances 0.000 description 2
- 239000004925 Acrylic resin Substances 0.000 description 1
- 229920000178 Acrylic resin Polymers 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000000806 elastomer Substances 0.000 description 1
- 230000002209 hydrophobic Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Description
【特許請求の範囲】
【請求項１】 化学反応を制御するカートリッジにおいて、このカートリッジは、
a) 内部に形成された少なくとも第１と第２の流路を有する本体と、
b) この本体から延び出す反応容器とを備え、この反応容器は、
i) 反応室と、
ii) この反応室に入口流路を介して接続される入口ポートと、
iii) 上記反応室に出口流路を介して接続される出口ポートとを有し、上記反応容器の入口ポートは、上記本体内の第１の流路に接続され、上記反応容器の出口ポートは、上記本体内の第２の流路に接続されているカートリッジ。
【請求項２】 請求項１に記載のカートリッジにおいて、上記本体は、上記第２の流路に連通してガスを逃がすベントを更に有するカートリッジ。
【請求項３】 請求項１または２に記載のカートリッジにおいて、上記本体の第１の流路内の流体を上記反応容器の入口ポートを経て上記反応室に送る差圧源を更に有するカートリッジ。
【請求項４】 請求項１乃至３のいずれか１つに記載のカートリッジにおいて、上記反応容器は、
i) 上記反応室の側壁を区画する剛な枠と、
ii) 上記反応室の主壁を形成すべく上記剛な枠に取り付けられる少なくとも１つの柔軟なシートまたは膜とを有し、上記主壁は、熱面に一致するように十分柔軟であるようなカートリッジ。
【請求項５】 請求項１乃至３のいずれか１つに記載のカートリッジにおいて、上記反応容器は、
i) 上記反応室の側壁を区画する剛な枠と、
ii) 上記反応室の対向する主壁を形成すべく上記剛な枠の対向する側面に取り付けられる第１と第２の柔軟なシートまたは膜とを有するカートリッジ。
【請求項６】 請求項５に記載のカートリッジにおいて、上記第１と第２の柔軟なシートまたは膜は、夫々ポリマー膜からなるカートリッジ。
【請求項７】 請求項４または５に記載のカートリッジにおいて、上記側壁の少なくとも２つは、透光性であるとともに、互いに角度的に偏らせられているカートリッジ。
【請求項８】 請求項１乃至７のいずれか１つに記載のカートリッジにおいて、上記反応室の厚さに対する幅の比は、少なくとも２:１であるカートリッジ。
【請求項９】 請求項１乃至８のいずれか１つに記載のカートリッジにおいて、上記反応室は、５mm以下の厚さを有するカートリッジ。
【請求項１０】 請求項１乃至９のいずれか１つに記載のカートリッジにおいて、上記反応室は、0.5〜５mmの範囲の厚さを有するカートリッジ。
【請求項１１】 請求項１乃至７のいずれか１つに記載のカートリッジにおいて、上記反応室の厚さに対する幅の比は、少なくとも４:１であるカートリッジ。
【請求項１２】 請求項１乃至７または請求項１１のいずれか１つに記載のカートリッジにおいて、上記反応室は、0.5〜２mmの範囲の厚さを有するカートリッジ。
【請求項１３】 請求項１乃至７または１１乃至１２のいずれか１つに記載のカートリッジにおいて、上記反応室は、２mm以下の範囲の厚さを有するカートリッジ。
【請求項１４】 請求項１乃至１３のいずれか１つに記載のカートリッジにおいて、上記本体は、上記反応容器の入口ポートに上記第１の流路を経て接続され、流体試料を増幅試薬と混合するための混合室を更に有するカートリッジ。
【請求項１５】 請求項１乃至１４のいずれか１つに記載のカートリッジにおいて、上記本体は、内部に形成された
i) 試料流路と、
ii) 試料流体から所望の分析対象物を分離すべく、上記第１の流路を経て上記入口ポートに接続される上記試料流路内の分離領域とを有するカートリッジ。
【請求項１６】 請求項１５に記載のカートリッジにおいて、上記本体内の分離領域は、
a) 上記試料流路に設けられ、上記試料中の細胞またはウイルスを溶解させて、これらの細胞またはウイルスから物質を解き放たせる溶解室と、
b) 上記溶解させられるべき細胞またはウイルスを捕捉すべく、上記溶解室内に設けられた少なくとも１つの固体の担体とを有するカートリッジ。
【請求項１７】 請求項１に記載のカートリッジを有する装置であって、上記反応容器は、上記反応室を区画する複数の壁を有し、少なくとも１つの壁は、柔軟なシートまたは膜からなり、上記装置は、
a) 上記シートまたは膜に接触する少なくとも１つの熱面と、
b) 上記反応室内の圧力の増加が、上記シートまたは膜を上記熱面に一致するように接触させるに十分であるように、上記反応室内の圧力を増大させる手段と、
c) 上記反応室内の温度変化を引き起こさせるべく上記熱面を加熱または冷却する少なくとも１つの熱要素とを更に有する装置。
【請求項１８】 請求項１乃至３のいずれか１つに記載のカートリッジにおいて、上記反応容器は、
i) ２つの対向する主壁と、
ii) 上記主壁を互いに連結して、上記反応室を区画する側壁とを備え、上記側壁の少なくとも２つは、透光性で互いに角度的に偏らせられているカートリッジ。
【請求項１９】 請求項１乃至３のいずれか１つに記載のカートリッジを有する装置であって、上記反応容器は、２つの対向する主壁と、上記主壁を互いに連結して、上記反応室を区画する側壁とを備え、上記側壁の少なくとも２つは、透光性であり、上記透光性の側壁のうちの第１の側壁を介して上記反応室内の反応混合物を励起する少なくとも１つの光源と、上記反応室から上記透光性の側壁のうちの第２の側壁を介して放出される光を検出する少なくとも１つの検出器とを有する光学システムを更に備えている装置。
【請求項２０】 請求項１乃至３または８乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応室の各側面は、１〜１５mmの長さを持つカートリッジ。
【請求項２１】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応容器は、上記反応室を通る光路長が、１〜１５mmであるカートリッジ。
【請求項２２】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応容器は、上記反応室を通る光路長が、５〜１２mmであるカートリッジ。
【請求項２３】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応室は、体積容量が１００μリッター以下であるカートリッジ。
【請求項２４】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応室は、体積容量が１２〜１００μリッターであるカートリッジ。
【請求項２５】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応室の壁の少なくとも１つは、0.5mm以下の厚さであるカートリッジ。
【請求項２６】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記反応室の壁の少なくとも１つは、0.003〜0.5mmの厚さであるカートリッジ。
【請求項２７】 請求項１乃至１６のいずれか１つに記載のカートリッジにおいて、上記本体内の第１と第２の流路の幅または直径は、0.4〜3.2mmであるカートリッジ。
[Claims]
1. A cartridge for controlling a chemical reaction, the cartridge comprising:
a) at least formed insideFirst and secondA body having a flow path;
b) a reaction vessel extending from the body, the reaction vessel comprising:
i)Reaction chamberWhen,
ii)Inlet port connected to this reaction chamber via an inlet channelWhen,
iii)Outlet port connected to the reaction chamber via an outlet channelAnd havingAn inlet port of the reaction vessel is connected to a first flow path in the main body, and an outlet port of the reaction vessel is connected to a second flow path in the main body.cartridge.
2. The cartridge according to claim 1, whereinThe main body further includes a vent communicating with the second flow path to release gas.Having a cartridge.
3. The method according to claim 1,Or 2In the cartridge described in the above,The apparatus further includes a differential pressure source that sends a fluid in the first flow path of the main body to the reaction chamber via an inlet port of the reaction vessel.cartridge.
Claim 4Any one of 1 to 3In the cartridge described in the above,The reaction vessel is
i) a rigid frame defining the side wall of the reaction chamber;
ii) at least one flexible sheet or membrane attached to the rigid frame to form a main wall of the reaction chamber, wherein the main wall is sufficiently flexible to conform to a hot surface.cartridge.
Claim 5Any one of 1 to 3In the cartridge described in the above,The reaction vessel is
i) a rigid frame defining the side wall of the reaction chamber;
ii) having first and second flexible sheets or membranes attached to opposing sides of the rigid frame to form opposing main walls of the reaction chambercartridge.
Claim 65In the cartridge described in the above,The first and second flexible sheets or membranes each comprise a polymer membranecartridge.
7. Claim4 or 5In the cartridge described in the above,At least two of the side walls are translucent and angularly offset from one another.cartridge.
Claim 8Any one of 1 to 7In the cartridge described,The ratio of the width to the thickness of the reaction chamber is at least 2: 1cartridge.
9. The method of claim 1,To any one of 8In the cartridge described,The reaction chamber has a thickness of 5 mm or lesscartridge.
Claim 10Any one of 1 to 9In the cartridge described,The reaction chamber has a thickness in the range of 0.5-5mmcartridge.
11. The method according to claim 1,To any one of 7In the cartridge described,The ratio of the width to the thickness of the reaction chamber is at least 4: 1cartridge.
Claim 12Any one of claims 1 to 7 or claim 11In the cartridge described,The reaction chamber has a thickness in the range of 0.5 to 2 mmcartridge.
Claim 131 to 7 or 11 to 12In the cartridge described,The reaction chamber has a thickness in the range of 2 mm or lesscartridge.
Claim 14Any one of 1 to 13In the cartridge described,The main body is further connected to the inlet port of the reaction vessel via the first flow path, and further has a mixing chamber for mixing the fluid sample with the amplification reagent.cartridge.
Claim 15Any one of 1 to 14In the cartridge described in the above,The body is formed inside
i) sample flow path;
ii) having a separation area in the sample flow path connected to the inlet port via the first flow path to separate a desired analyte from the sample fluid.cartridge.
Claim 16FifteenIn the cartridge described in the above,The separation area in the main body is
a) a lysis chamber provided in the sample flow path to lyse cells or viruses in the sample and release substances from these cells or viruses;
b) a cartridge having at least one solid carrier provided in the lysis chamber to capture the cells or viruses to be lysed.
17. An apparatus having the cartridge according to claim 1, wherein the reaction container has a plurality of walls defining the reaction chamber, at least one wall is formed of a flexible sheet or a membrane, and the apparatus includes:
a) at least one hot surface in contact with said sheet or membrane;
b) means for increasing the pressure in the reaction chamber, such that the increase in pressure in the reaction chamber is sufficient to bring the sheet or membrane into contact with the hot surface;
c) at least one heating element for heating or cooling the hot surface to cause a temperature change in the reaction chamber..
18. The method of claim 1,Any one of 3In the cartridge described in the above,The reaction vessel is
i) two opposing main walls;
ii) a side wall connecting the main walls to each other to partition the reaction chamber, at least two of the side walls being translucent and angularly offset from each other.cartridge.
(19) Any one of claims 1 to 3Cartridge described inWherein the reaction vessel includes two opposing main walls, and a side wall connecting the main walls to each other to define the reaction chamber, and at least two of the side walls are light-transmitting. At least one light source that excites a reaction mixture in the reaction chamber through a first one of the translucent sidewalls, and a second one of the translucent sidewalls from the reaction chamber. Further comprising an optical system having at least one detector for detecting light emitted through a side wall of the device..
20. Any one of claims 1 to 3 or 8 to 16In the cartridge described in the above,Each side of the reaction chamber has a length of 1 to 15 mmcartridge.
21. Any one of claims 1 to 16In the cartridge described in the above,The reaction vessel has an optical path length of 1 to 15 mm passing through the reaction chamber.cartridge.
22. The cartridge according to any one of claims 1 to 16, wherein the reaction vessel has an optical path length passing through the reaction chamber of 5 to 12 mm..
23. The cartridge according to any one of claims 1 to 16, wherein the reaction chamber has a volume capacity of 100 µl or less.
24. The cartridge according to any one of claims 1 to 16, wherein the reaction chamber has a volume capacity of 12 to 100 µl.
25. 17. The cartridge according to claim 1, wherein at least one of the walls of the reaction chamber has a thickness of 0.5 mm or less..
26. 17. The cartridge according to any one of claims 1 to 16, wherein at least one of the walls of the reaction chamber has a thickness of 0.003 to 0.5 mm..
27. The cartridge according to any one of claims 1 to 16, wherein the width or diameter of the first and second flow paths in the main body is 0.4 to 3.2 mm..
図１は、好ましい実施形態のカートリッジ２０の等角投影図である。カートリッジ２０は、流体試料から核酸を分離するように設計されており、増幅および検出のために核酸を保持するように設計されている。カートリッジ２０は、頂部片２２,中央片２４および底部片２６を備える本体を有している。流体試料をカートリッジの中に導入するための入口は頂部片２２に形成されてキャップ３０によって密閉されている。また、６つの圧力ポート３２が、頂部片２２に形成されている。圧力ポート３２は、例えばポンプや真空などの圧力源からのノズルを収容するためにある。カートリッジはまた、装置(図１０で後述する)におけるカートリッジ２０の位置決めのために底部片２６から延び出る整列脚２８を有する。窪みまたは凹部３８A,３８B,３８Cは、頂部片２２および中央片２２に形成されている。この凹部は、カートリッジ２０内の流体の流れを検出するための光学センサを収容するためにある。カートリッジ２０は、更にベント３４,３６を有する。各圧力ポートと各ベントは、ベントおよび圧力ポートに対してガスが出入りするようにガスは通すが、液体は通さない疎水性の薄膜を有することが好ましい。変性アクリル樹脂共重合体の薄膜は、例えば、ゲルマンサイエンス社(ミシガン州,アン・アーバー)から市販され、また、粒子トラック腐食ポリカーボネートの薄膜は、ポリティクス社(カリフォルニア州,リバーモア)から市販されている。 FIG. 1 is an isometric view of the cartridge 20 of the preferred embodiment. Cartridge 20 is designed to separate nucleic acids from a fluid sample and is designed to retain nucleic acids for amplification and detection. The cartridge 20 has a body with a top piece 22, a center piece 24 and a bottom piece 26. An inlet for introducing a fluid sample into the cartridge is formed in the top piece 22 and is closed by a cap 30. Also, six pressure ports 32 are formed in the top piece 22. The pressure port 32 is for receiving a nozzle from a pressure source such as a pump or vacuum. The cartridge also has alignment feet 28 extending from the bottom piece 26 for positioning of the cartridge 20 in the device (described below in FIG. 10). Recesses or recesses 38A, 38B, 38C are formed in the top piece 22 and the center piece 22. The recess is provided for accommodating an optical sensor for detecting the flow of the fluid in the cartridge 20. The cartridge 20 further has vents 34 and 36. Each pressure port and each vent preferably has a hydrophobic thin film that allows gas to pass through but does not allow liquid to enter and exit the vent and pressure port. Thin films of modified acrylic resin copolymers are commercially available, for example, from Germanic Science Inc. I have.
図５を再度参照すると、溶解室８６の加圧は、トランスデューサ９２と溶解室８６の可撓性壁６３との間の結合を確実かつ有効なものにするので重要である。溶解室８６内の細胞あるいはウイルスを崩壊させるために、トランスデューサ９２が作動される(すなわち振動運動するようにセットされる)。溶解室８６の可撓性壁６３は、僅かな偏向によって、壁内に高圧を生じることなくトランスデューサ９２の振動を室８６内の液体に伝える。上述したように、壁６３はエラストマの膜によって形成される。この代わりに、壁は、好ましくは0.025〜0.1mmの範囲にある厚みのポリマー材の膜またはシート(例えばプリプロピレン膜)である。トランスデューサ９２は、好ましくは、室８６を超音波処理するための超音波ホーンである。室８６は、好ましくは、２０〜６０ｋＨｚの範囲にある周波数で１０〜４０秒間超音波処理される。例示のプロトコルでは、室は４７ｋＨｚの周波数で１５秒間超音波処理される。ホーンチップの振幅は、好ましくは、２０〜２５μｍの範囲にある(ピーク間で測定)。 Referring again to FIG. 5, pressurization of the lysis chamber 86 is important because it ensures a secure and effective connection between the transducer 92 and the flexible wall 63 of the lysis chamber 86. To disrupt the cells or c i Luz dissolution chamber 86, the transducer 92 (which is set to That vibratory motion) that is being operated. The flexible wall 63 of the lysis chamber 86 transmits the vibration of the transducer 92 to the liquid in the chamber 86 with a slight deflection without creating high pressure in the wall. As described above, the wall 63 is formed by an elastomer film. Alternatively, the wall is preferably film or sheet of polymer over material thickness in the range of 0.025～0.1Mm (e.g. prepro Hoon Renmaku). Transducer 92 is preferably an ultrasonic horn for sonicating chamber 86. The chamber 86 is preferably sonicated at a frequency in the range of 20-60 kHz for 10-40 seconds. In the exemplary protocol, the chamber is sonicated for 15 seconds at a frequency of 47 kHz. The horn tip amplitude is preferably in the range of 20-25 μm (measured between peaks).
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13670399P | 1999-05-28 | 1999-05-28 | |
| US60/136,703 | 1999-05-28 | ||
| US09/469,724 US6431476B1 (en) | 1999-12-21 | 1999-12-21 | Apparatus and method for rapid ultrasonic disruption of cells or viruses |
| US09/469,724 | 1999-12-21 | ||
| PCT/US2000/014739 WO2000072970A1 (en) | 1999-05-28 | 2000-05-30 | Cartridge for conducting a chemical reaction |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2009180791A Division JP4800412B2 (en) | 1999-05-28 | 2009-08-03 | Method for conducting chemical reaction in the cartridge |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2003500674A JP2003500674A (en) | 2003-01-07 |
| JP2003500674A5 true JP2003500674A5 (en) | 2006-12-21 |
| JP4495866B2 JP4495866B2 (en) | 2010-07-07 |
Family
ID=26834561
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2000621073A Expired - Lifetime JP4495866B2 (en) | 1999-05-28 | 2000-05-30 | Cartridge for controlling chemical reactions |
| JP2009180791A Expired - Lifetime JP4800412B2 (en) | 1999-05-28 | 2009-08-03 | Method for conducting chemical reaction in the cartridge |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2009180791A Expired - Lifetime JP4800412B2 (en) | 1999-05-28 | 2009-08-03 | Method for conducting chemical reaction in the cartridge |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP1181098B2 (en) |
| JP (2) | JP4495866B2 (en) |
| AT (1) | ATE333938T1 (en) |
| AU (1) | AU781122B2 (en) |
| CA (1) | CA2373198C (en) |
| DE (1) | DE60029582T3 (en) |
| DK (1) | DK1181098T4 (en) |
| ES (1) | ES2272289T5 (en) |
| WO (1) | WO2000072970A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US11845081B2 (en) | 2007-07-13 | 2023-12-19 | Handylab, Inc. | Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples |
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