JP6061313B2 - 核酸を処理及び検出するためのマイクロ流体カートリッジ - Google Patents
核酸を処理及び検出するためのマイクロ流体カートリッジ Download PDFInfo
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- JP6061313B2 JP6061313B2 JP2014556810A JP2014556810A JP6061313B2 JP 6061313 B2 JP6061313 B2 JP 6061313B2 JP 2014556810 A JP2014556810 A JP 2014556810A JP 2014556810 A JP2014556810 A JP 2014556810A JP 6061313 B2 JP6061313 B2 JP 6061313B2
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Description
図1A〜図1Cに示すように、核酸を処理及び検出するためのマイクロ流体カートリッジ100の一実施形態は、一連のサンプルポート−試薬ポートの対112及び一連の検出チャンバ116を備える最上層110と、最上層110に結合されて、廃棄チャンバ130を形成するように構成されたフィルム層125によって最上層から部分的に隔てられた中間基板120と、中間基板120上に部分的に位置する弾性層140と、磁界156を提供する磁石152がアクセス可能な磁石ハウジング領域150と、各々が、少なくとも、最上層110の一部、フィルム層125の一部及び弾性層140の一部によって形成された一連の流路160と、を備えている。他の実施形態では、マイクロ流体カートリッジ100は、中間基板120に結合されて廃棄チャンバ130を密閉するように構成された最下層170をさらに備えてもよい。さらに、マイクロ流体カートリッジ100の最上層110は、共有流体ポート118と、排出領域190と、加熱領域195と、をさらに備えてもよく、一連の流路160の各流路165が、各サンプルポート−試薬ポートの対113、共有流体ポート118、廃棄チャンバ130及び検出チャンバ117に流体的に結合され、各流路165は、加熱領域及び磁界を通過するように構成された捕捉区画166を備えており、かつ、検出チャンバ117の上流で排出領域190を通過するように構成されている。従って、各流路165は、核酸を含んだサンプル流体が流路165の様々な部分を通過する際に、サンプル流体を受け入れてサンプル流体の処理を促進するように機能する。そのように構成されるので、マイクロ流体カートリッジ100は、分子診断処理及び技術を促進するように用いられることが可能であり、かつ、マイクロタイタープレートの寸法基準に適合することが好ましい。代替として、マイクロ流体カートリッジ100は任意の適切なサイズであってもよい。特定の用途において、マイクロ流体カートリッジ100は、核酸を含んだサンプルの分析のためのPCR処理を促進するために用いることが可能である。
一実施形態のマイクロ流体カートリッジ100の最上層110は、カートリッジを通り抜ける核酸を含んだサンプルが、分子診断処理(例えばPCR)の実行に関与するエレメントによって操作されることが可能なように、分子診断処理の実行に関与するエレメントを収容するように機能する。最上層110は、当該最上層110が、蛍光技術又は化学発光技術によるサンプルの検出、適切なガラス転移温度、及び、PCRに対する化学的適合性、又は他の増幅技術と干渉しないように、低い自己蛍光性を有する構造的に剛体の/堅固な材料から形成されることが好ましい。好ましくは、最上層110は、ポリプロピレンベースのポリマーから形成されるものの、最上層110は、代替として、任意の適切な材料(例えば環状オレフィンポリマー)から形成されてもよい。特定の実施形態では、最上層110は、136〜163℃のガラス転移温度を有するとともに射出成形によって製造された1.5mmの厚さのポリプロピレンから形成される。最上層110は、代替として、例えばポリプロピレンベースのポリマーといった任意の適切な材料から形成されてもよい。図1B及び図1Cに示すように、最上層110は、一連のサンプルポート−試薬ポートの対112と、流体ポート118と、排出領域190と、流路165の捕捉区画166を横切る加熱領域195と、一連の検出チャンバ116と、を備えることが好ましい。
図1Bに示すように、一実施形態のマイクロ流体カートリッジは、最上層110に結合された中間基板120であって、廃棄チャンバ130を形成するように構成されたフィルム層125によって最上層110から部分的に隔離された中間基板120を備えている。中間基板120は、バルブピンのためのガイドを提供するため、及び、廃棄流体が堆積し得る廃棄チャンバボリュームを提供するため、マイクロ流体カートリッジの層が接合され得る基板としての役割を果たすように機能する。好ましくは、中間基板120の深さは、マイクロ流体カートリッジ100内で生成された廃棄流体のボリュームを収容するのに十分な廃棄チャンバボリュームを提供する。さらに、中間基板120の深さは、小型の分子診断システムのすべての場所でのマイクロ流体カートリッジ100の移動を容易にするために薄型である。好ましくは、マイクロ流体カートリッジ100の中間基板120は、マイクロ流体カートリッジ100の自動操作を容易にするため、マイクロタイタープレート標準を遵守するように構成される。中間基板120は、例えばポリプロピレンなどの低コストで構造的に堅固な材料から構成されることが好ましい。しかしながら、最上層120と同様に、中間基板は、代替として、当該中間基板120が、蛍光技術によるサンプル検出と、PCR技術のための適切なガラス転移温度と干渉しないように、低い自己蛍光性を有する構造的に堅固な材料から形成されてもよい。この代替実施形態の一変形例では、中間基板120は、136〜163℃のガラス転移温度を有する、射出成形によって製造された環状オレフィンポリマー(COP)から構成される。さらに別の代替実施形態では、中間基板120は、任意の適切な材料、例えばポリカーボネートベースのポリマーから構成されてもよい。
図1B及び図5A〜図5Dに示すように、一実施形態のマイクロ流体カートリッジ100はまた、中間基板120上に部分的に位置する弾性層140を備えており、弾性層140は、変形時、一連の閉塞位置141のうちの1つの閉塞位置で弾性層140に接触する流路165のマイクロ流体チャネルを閉塞する変形可能基板を提供するように機能する。好ましくは、弾性層140は、適切な厚さを有する不活性の液体不透過性材料であってよく、この材料は、製造中に直面する温度、及び/又は、分子診断プロトコルで特定される温度まで、実質的に損傷(例えば傷つけられた表面及び/又は機械的なロバスト性の欠落)がない状態で、加熱されることが可能であり、PCRアッセイと化学的に適合する。好ましくは、弾性層140は非連続的であり、その結果、弾性層140の一部が、一連のバルブガイド127によって提供された孔を直接覆うように中間基板120に対して位置決めされる。代替として、弾性層140は、マイクロ流体カートリッジ100の設置面積の大部分にわたって広がる連続的な層であり、一連のバルブガイド127によって提供された孔を覆う。特定の実施形態では、弾性層140は、実質的な損傷なく少なくとも120℃まで加熱可能な低デュロメータのシリコーンの500ミクロンの厚さのストリップを備えており、ストリップは、シリコーンベースの接着剤を用いて中間基板120の一部に接合されて、フィルム層125及び中間基板120の間でわずかに圧縮される。特定の実施形態の一変形例では、弾性層140は代替として、中間基板120と最上層110との間の圧力のみによって所定の位置に保持されてもよい。好ましくは、弾性層140は、マイクロ流体カートリッジ100の使用寿命にわたって可逆的に変形可能であり、その結果、弾性層140に接触する流路165のマイクロ流体チャネルの任意の閉塞は、マイクロ流体カートリッジの使用寿命にわたって可逆的である。代替として、弾性層140は、可逆的に変形可能でなくてもよく、その結果、弾性層140に接触する流路165のマイクロ流体チャネルの閉塞は可逆的ではない。
マイクロ流体カートリッジ100の磁石ハウジング領域150は、核酸の精製及び分離のために磁界156を提供する少なくとも1つの磁石152に対するアクセスを提供する及び/又は磁石152を収容するように機能する。好ましくは、磁石ハウジング領域150はフィルム層及び中間基板によって規定され、その結果、フィルム層及び中間基板は磁石ハウジング領域150の境界を形成する。最下層170を備えるマイクロ流体カートリッジ100の一実施形態では、磁石ハウジング領域150は、最下層170によってさらに規定されてもよく、その結果、最下層は、磁石ハウジング領域150の境界を部分的に形成する。磁石ハウジング領域150は、マイクロ流体カートリッジ150の矩形のプリズム形状の空隙であることが好ましく、かつ、図1Bに示すように、マイクロ流体カートリッジ100の一面を通じてのみアクセス可能であることが好ましい。好ましくは、磁石ハウジング領域150は、磁石152を収容するために磁石152上を可逆的に通過させられることが可能であり、磁石ハウジング領域150から磁石152を取り外すために後退させられることが可能であるが、磁石152は、磁石152が磁石ハウジング領域150に進入すると、代替として、磁石ハウジング領域150内に不可逆的に固定されてもよい。
マイクロ流体カートリッジ100の一連の流路160は、分子診断プロトコルで用いられる多量のサンプル流体、試薬、バッファ及び/又は気体が送達され得る流体ネットワークを提供し、一連の流路160から外側に廃棄流体が排出され、かつ、一連の流路160によって、処理された核酸サンプルが、増幅及び/又は検出を含み得る分析のための検出チャンバに送達され得る。好ましくは、一連の流路160の各流路165は、少なくとも、最上層の一部、フィルム層の一部及び弾性層140の一部によって形成されており、その結果、各流路165は、一連の閉塞位置141の弾性層140の変形時に閉塞され得る。さらに、一連の流路160の少なくとも1つの流路165は、一連のサンプルポート−試薬ポートの対112のサンプルポート−試薬ポートの対113、流体ポート118、廃棄チャンバ130及び一連の検出チャンバ116の検出チャンバ117に流体的に結合されることが好ましい。さらに、一連の流路160の少なくとも1つの流路165は、弾性層140の変形時に閉塞されるように構成され、廃棄チャンバ30に廃棄流体を運搬するように構成され、加熱領域195及び磁界156を通過する捕捉区画166を備え、かつ、検出チャンバ117の上流で排出領域190を通過するように構成されることが好ましい。代替実施形態は、例えば排出領域190又は加熱領域195などの上述の流路165の実施形態の好適なエレメントを省略してもよく、又は、上述の流路165の実施形態にさらなるエレメントを追加してもよい。
マイクロ流体カートリッジ100は、実際のバルブ部材がマイクロ流体カートリッジ100に一体化されないように、従って、流路165の開放部分及び/又は閉塞部分が、マイクロ流体カートリッジの外側に配置されたシステムによって実行されるように、構成されることが好ましい。一例として、流路165の一部が、上述したように、弾性層140を変形させて流路165を閉塞するためのバイアス力を付与する、カードの下に配置されたバルブ部材又はバルブ機構の動作によって、閉塞位置で開放され又は閉塞されてよい。力は、図10A及び図10Bに示すように、機械的部材(例えばピン、ポストなど)、電気機械部材(例えばソレノイド)、空気圧式部材若しくは水圧式部材(例えば空気、水など)、又は、任意の他の適切な手段によって付与されてもよい。ある変形例では、カートリッジは、バルブ部材又はバルブ機構に対してカードが整列させられることを可能にする1以上の登録領域を含んでもよい。代替実施形態では、弾性層140、一連のバルブガイド127、及び、一連の閉塞位置141は、省略され、流路165の部分を制御可能に閉塞して開放するように構成された、マイクロ流体カートリッジ100内に一体化されたバルブと置換されてもよい。
マイクロ流体カートリッジ100の特定の実施形態に関する以下の説明は、例示のみの目的のためになされ、請求項に係る発明の範囲の確定的なものとして又は制限するものとして解釈されるべきではない。
マイクロ流体カートリッジ100の実施形態のための組立方法200の実施形態を図12A〜図12Gに示す。組立方法200は、フィルム層に最上層を整列させてその2つを熱接合し、シリコーン接着剤を用いてマイクロ流体カートリッジの中間基板に弾性層を接合するステップS210と、最上層、フィルム層、弾性層及び中間基板を圧縮して、最上層/フィルム層を弾性層/中間基板に接合するステップS220と、最下層に中間基板を接合するステップS230と、排出領域の排出口を設置するステップS250と、ラベルを貼って包装するステップS260と、を含むことが好ましい。
Claims (14)
- 核酸の処理及び検出を促進するように構成されたカートリッジであって、
サンプルポート、試薬ポート、流体ポート及び検出チャンバを有する第1層と、
波形表面を有する廃棄チャンバを規定する中間基板と、
弾性層と、
少なくとも前記第1層の一部と前記弾性層の一部とによって形成された流路であって、前記サンプルポート、前記試薬ポート、前記流体ポート及び前記検出チャンバに流体的に結合されており、かつ、前記弾性層の操作時に一連の閉塞位置で閉塞されるように構成された流路と、を備え、
前記一連の閉塞位置の第1サブセットでの前記流路の操作時に、第1の先を断ち切った通路が規定され、
前記一連の閉塞位置の第2サブセットでの前記流路の操作時に、前記検出チャンバまでの第2の先を断ち切った通路が規定される、カートリッジ。 - 前記第1層は、液体不透過性膜を有する排出領域をさらに有しており、前記流路は、前記検出チャンバの上流で前記排出領域を通過するように構成される、請求項1に記載のカートリッジ。
- 前記中間基板が、前記第1層に結合されて、一連のバルブガイドを規定し、前記波形表面は、前記第1の先を断ち切った通路内で電磁ビーズに結合された核酸を捕捉するように構成された磁界を提供する磁石を受け入れるように構成された領域を規定する、請求項1に記載のカートリッジ。
- 前記一連の閉塞位置の第3サブセットで枝分かれした前記流路の閉塞時に、前記流体ポートに結合される一方で前記サンプルポート及び前記試薬ポートには結合されない第3の先を断ち切った通路が規定され、前記一連の閉塞位置の前記第1サブセット、前記一連の閉塞位置の前記第2サブセット及び前記一連の閉塞位置の前記第3サブセットは重なっており、かつ、前記一連の閉塞位置は、少なくとも1つの通常開放位置と1つの通常閉鎖位置とを備える、請求項1に記載のカートリッジ。
- 核酸の処理を促進するように構成されたカートリッジであって、
サンプルポート、試薬ポート及び流体ポートと、
磁石を受け入れるように構成された磁石領域と、
前記サンプルポート、前記試薬ポート及び前記流体ポートに流体的に結合された流路であって、前記磁石領域を横切るように構成された捕捉区画を備え、前記捕捉区画は、前記磁石領域を横切るように構成された転換区画であって、前記磁石によって提供された磁界によって電磁ビーズに結合された核酸を捕捉するように構成された転換区画である、流路と、
第1層、中間基板及び弾性層とを備え、
前記第1層は、前記サンプルポート、前記試薬ポート、前記流体ポート及び検出チャンバを規定し、
前記中間基板は、前記弾性層が前記中間基板と前記第1層との間に位置するように前記第1層に結合するように構成され、かつ、前記磁石領域を規定し、
前記流路は、少なくとも前記第1層の一部と前記弾性層の一部とによって形成され、前記弾性層の操作時に閉塞されるように構成されており、
前記中間基板は、フィルム層によって前記第1層から部分的に分離され、前記中間基板は、前記磁石領域、前記流路に結合された廃棄物入口、及び、廃棄物排出口を部分的に形成するように構成された廃棄チャンバをさらに規定する、カートリッジ。 - 排出領域及び加熱領域をさらに備え、前記流路の前記捕捉区画は、前記磁石領域及び前記加熱領域を横切るように構成され、前記流路は、前記検出チャンバの上流で前記排出領域を通過するように構成される、請求項5に記載のカートリッジ。
- 核酸の処理及び検出を促進するように構成されたカートリッジであって、
第1層と、前記第1層に結合されて波形表面を有する廃棄チャンバを規定する中間基板と、
少なくとも前記第1層の一部によって形成された第1流路と、
少なくとも前記第1層の一部によって形成された第2流路であって、前記第1流路は前記廃棄チャンバに廃棄物を伝達するように構成され、前記第2流路は前記廃棄チャンバに廃棄物を伝達するように構成される、第2流路と、を備えるカートリッジ。 - 前記第1層は、第1サンプルポート−試薬ポートの対、第2サンプルポート−試薬ポートの対、共有流体ポート、第1検出チャンバ、及び、第2検出チャンバを備え、前記第1流路は、前記第1サンプルポート−試薬ポートの対と前記第1検出チャンバとに結合され、前記第2流路は、前記第2サンプルポート−試薬ポートの対と前記第2検出チャンバとに結合され、前記共有流体ポートは、前記第1流路及び前記第2流路に流体的に結合するように構成される、請求項7に記載のカートリッジ。
- 加熱領域及び排出領域をさらに備え、前記第1流路は、前記加熱領域を横切って前記第1検出チャンバの上流で前記排出領域を通過するように構成され、前記第2流路は、前記加熱領域を横切って前記第2検出チャンバの上流で前記排出領域を通過するように構成され、前記加熱領域は、前記カートリッジの凹部領域によって規定されて前記カートリッジの長さにわたって広がる、請求項8に記載のカートリッジ。
- 前記排出領域は、気体透過性である疎水性の液体不透過性の膜を備える、請求項6又は9に記載のカートリッジ。
- 前記第1検出チャンバは第1蛇行形状流路を備え、前記第2検出チャンバは第2蛇行形状流路を備え、前記第1蛇行形状流路及び前記第2蛇行形状流路のうちの少なくとも1つが、2つの狭チャネルによって相互に結合される3つの広チャネルを備える、請求項8に記載のカートリッジ。
- 前記第1検出チャンバ及び前記第2検出チャンバは同一であり、前記第1検出チャンバは片側から加熱されるように構成され、前記第2検出チャンバは片側から加熱されるように構成され、前記第1検出チャンバ及び前記第2検出チャンバのうちの少なくとも1つが、気泡発生を制限するように、容積、熱サイクリング率、光検出及び充填に関して最適化されるように構成される、請求項8に記載のカートリッジ。
- 波形表面が磁石ハウジング領域を規定し、前記廃棄チャンバが前記第1層の下方に配置される、請求項7に記載のカートリッジ。
- 前記カートリッジはマイクロタイタープレートの寸法基準に従う、請求項1、5及び7のいずれか1項に記載のカートリッジ。
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US11213824B2 (en) | 2017-03-29 | 2022-01-04 | The Research Foundation For The State University Of New York | Microfluidic device and methods |
US11911763B2 (en) | 2017-03-29 | 2024-02-27 | The Research Foundation For The State University Of New York | Microfluidic device and methods |
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