JP2017513496A - 細胞の検出のための試薬カートリッジ及び方法 - Google Patents
細胞の検出のための試薬カートリッジ及び方法 Download PDFInfo
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Abstract
Description
本願は、2015年2月9日に出願された「Reagent Cartridge and Methods for Detection of Cells」と題する米国特許出願第14/617,631号明細書の継続であり、それに基づく優先権を主張する。該出願は、2014年9月3日に出願された「Reagent Cartridge and Methods for Detection of Cells」と題する米国特許出願第14/476,392号明細書の一部継続であり、2014年4月24日に出願された「Reagent Cartridge for Detection of Cells」と題する米国仮特許出願第61/983,765号、2014年2月12日に出願された「Systems and Methods for Packaging Nucleic Acid Molecules into Non-Replicative Transduction Particles and Their Use as Cellular Reporters」と題する同第61/939,126号、及び2013年10月29日に出願された「Transcript Detection Systems and Methods」と題する同第61/897,040号に基づく優先権を主張する。該出願は各々、その内容全体を参照により本明細書に援用される。
Claims (47)
- 装置であって、
反応チャンバーに取り外し可能に連結されるように構成されたハウジングであって、試薬容器を受けるように構成される試薬ボリュームを画定し、前記試薬ボリュームと流体連通して転送経路を画定する穿孔器を含み、前記反応チャンバーに連結されるとき、前記転送経路と前記反応チャンバーの間に輸送経路を画定する輸送部分を含むハウジング;及び
前記試薬ボリューム内に配されるプランジャー部分を有するアクチュエーターであって、前記アクチュエーターの係合部分が、操作されて前記プランジャー部分を前記試薬ボリューム内で動かして前記試薬容器を変形させるように構成されたアクチュエーターを備え、前記穿孔器が、前記試薬容器の破壊可能部分を穿孔して試薬を前記試薬容器から前記反応チャンバーに前記転送経路及び前記輸送経路を介して運ぶように構成される装置。 - 前記転送経路が前記試薬容器の前記破壊可能部分に対して垂直である請求項1に記載の装置。
- 前記穿孔器が前記試薬ボリューム及び前記輸送経路と流体連通して複数の転送経路を画定し、前記転送経路が前記複数の転送経路の1つである請求項1に記載の装置。
- 前記穿孔器及び前記ハウジングが構造的に一体となって構築される請求項1に記載の装置。
- 前記輸送経路の部分が前記穿孔器を部分的に囲む請求項1に記載の装置。
- 前記輸送経路の出口部分が前記反応チャンバーの側壁と交わる出口軸線を画定する請求項1に記載の装置。
- 前記試薬ボリューム内に配され、前記破壊可能部分を囲む裾部を含み、前記裾部及び前記輸送部分の面が実質的に液密な封止を形成する前記試薬容器、並びに
前記試薬容器が変形して前記裾部と前記面の間の前記実質的に液密な封止を維持するとき、前記ハウジングに対する前記裾部の動きを制限するように構成される錠部材をさらに備える、請求項1に記載の装置。 - 前記試薬ボリューム内に配される前記試薬容器をさらに備え、前記試薬容器が前記破壊可能部分を囲む裾部を含み、前記裾部の第1の部分及び前記輸送部分の第1の面が、実質的に液密な封止を形成し、前記試薬容器が変形するとき、前記裾部の第2の部分が前記ハウジングの保持部分を係合して前記ハウジングに対する前記裾部の動き制限するように構成される、請求項1に記載の装置。
- 前記アクチュエーターの前記プランジャー部分及び前記ハウジングの部分が集合的に封止を画定して、前記試薬ボリュームを前記ハウジング外部のボリュームから流体的に隔てる請求項1に記載の装置。
- 前記試薬ボリューム内に配される前記試薬容器をさらに備え、前記試薬容器が複数の形質導入粒子を含有する、請求項1に記載の装置。
- 前記複数の形質導入粒子が溶解性複製不能であるように改変される請求項10に記載の装置。
- 前記試薬ボリューム内に配される前記試薬容器をさらに備え、前記試薬容器が試料中の複数のレポーター分子と反応して信号の生成を増強するように作成される試薬を含有する、請求項1に記載の装置。
- 前記試薬ボリューム内に配される前記試薬容器をさらに備え、前記試薬容器が少なくとも1つの栄養素、抗生物質、溶解試薬、又は滅菌試薬を含有する、請求項1に記載の装置。
- 前記試薬ボリューム内に配される前記試薬容器をさらに備え、前記試薬容器がトリデカナールを含有する、請求項1に記載の装置。
- 先細り状の側壁をもつ前記反応チャンバーをさらに備え、
前記輸送経路の出口部分が、前記反応チャンバーの前記先細り状の側壁と交わる出口軸線を画定する、請求項1に記載の装置。 - 装置であって、
反応チャンバーに取り外し可能に連結されるように構成されたハウジングであって、試薬ボリュームを画定し、前記反応チャンバーに連結されたとき、前記反応チャンバーと流体連通して前記試薬ボリュームを置く輸送経路を画定する、前記輸送経路と流体連通して穿孔器を含む輸送部分を含むハウジング;
前記ハウジングの前記試薬ボリューム内に配され、試薬を含有し、破壊可能部分及び前記破壊可能部分を囲む裾部を含む試薬容器;
前記試薬ボリューム内に配されるプランジャー部分を有するアクチュエーターであって、操作されて前記プランジャー部分を前記試薬ボリューム内で動かして前記試薬容器を第1の構成から第2の構成に変形させるように構成され、前記試薬容器が前記第2の構成にあるとき、前記穿孔器が、前記試薬容器の前記破壊可能部分を穿孔して試薬を前記試薬容器から前記反応チャンバーに前記輸送経路を介して運ぶように構成されるアクチュエーター;並びに
前記試薬容器が前記第2の構成にあるとき、前記裾部を前記ハウジングの前記輸送部分の肩と接触して維持して、前記裾部と前記肩の間の実質的に液密な封止を維持するように構成される錠部材を備える装置。 - 前記穿孔器が、前記試薬容器が第2の構成にあるとき、前記輸送経路と流体連通して前記反応チャンバーを置くように構成される複数の転送経路を画定する請求項16に記載の装置。
- 前記輸送経路の部分が前記穿孔器を部分的に囲む請求項16に記載の装置。
- 先細り状の側壁を有する前記反応チャンバーをさらに備え、
前記輸送経路の出口部分が前記反応チャンバーの前記先細り状の側壁と交わる出口軸線を画定する、請求項16に記載の装置。 - 前記第2の構成の前記試薬容器の容積が、前記第1の構成の前記試薬容器の容積の約5パーセント未満である請求項16に記載の装置。
- 前記試薬容器が複数の形質導入粒子を含有する請求項16に記載の装置。
- 前記試薬容器が、試料中の複数のレポーター分子と反応して信号の生成を増強するように作成された試薬を含有する請求項16に記載の装置。
- 装置であって、
反応チャンバーに取り外し可能に連結されるように構成されたハウジングであって、試薬ボリュームを画定し、前記反応チャンバーに連結されたとき、前記反応チャンバーと流体連通して前記試薬ボリュームを置く輸送経路を画定する、前記輸送経路と流体連通して穿孔器を含む輸送部分を含むハウジング;
前記ハウジングの前記試薬ボリューム内に配され、試薬を含有し、接触部分、破壊可能部分、及び前記破壊可能部分を囲む裾部を含む試薬容器;並びに
前記試薬容器又は前記穿孔器の少なくとも1つの前記接触部分に対応する、前記試薬ボリューム内に配されるプランジャー部分を有し、操作されて前記プランジャー部分を前記試薬ボリューム内で動かして、その結果前記プランジャー部分が前記試薬容器の前記接触部分に係合して前記試薬容器を第1の構成から第2の構成に変形させるように構成され、前記試薬容器が前記第2の構成にあるとき、前記穿孔器が、前記試薬容器の前記破壊可能部分を穿孔して試薬を前記試薬容器から前記反応チャンバーに前記輸送経路を介して運ぶアクチュエーターを備える装置。 - 前記試薬容器の前記接触部分が前記穿孔器を囲み、且つ
前記アクチュエーターの前記プランジャー部分が前記穿孔器と並んだ開口部を画定する、請求項23に記載の装置。 - 前記アクチュエーターの前記プランジャー部分の形状が、前記試薬容器の前記接触部分の形状に対応する、請求項20に記載の装置。
- 前記プランジャー部分が、前記試薬容器の前記接触部分の曲がった形状と嵌合して係合するように構成される曲がった形状を有する請求項23に記載の装置。
- 前記輸送部分が面及び肩を含み、前記穿孔器が前記面から伸び、前記試薬容器が前記第2の構成にあるとき、前記面が少なくとも前記試薬容器の前記破壊可能部分を受けるように構成され、前記肩が前記面を囲む、請求項23に記載の装置。
- 前記試薬容器が第2の構成にあるとき、前記裾部を前記ハウジングの前記輸送部分の前記肩と接触して維持して、前記裾部と前記肩の間の実質的に液密な封止を維持するように構成される錠部材をさらに備える、請求項23に記載の装置。
- 方法であって、
試料を、前記試料と混ぜてアッセイ培地を形成するように作成された試薬を含有するようにパッケージされた反応チャンバーに配すること;
前記反応チャンバー中で前記試料と、細胞表現型と関連する、前記細胞表現型に検出可能な信号を生成できる複数のレポーター分子を作らせるように作成された核酸分子を含むように改変された複数の形質導入粒子を混ぜることであって、前記試薬が前記細胞表現型の部分において検出可能な信号を抑制するように作成すること;
前記試薬と関連する第1の信号を受信すること;
前記第1の信号が前記試薬の存在を示す場合、前記試料及び前記複数の形質導入粒子を維持して、前記細胞表現型が前記試料中に存在するとき、前記複数のレポーター分子を発現させること;並びに
前記複数のレポーター分子の量と関連する第2の信号を受信することを含む方法。 - 前記試薬が乾燥試薬である請求項29に記載の方法。
- 前記試薬が前記反応チャンバーの内面に付着する請求項29に記載の方法。
- 前記細胞表現型の前記部分が抗生物質個々に又は別の抗生物質と組み合わせて耐性がある細菌表現型を含む請求項29に記載の方法。
- 前記抗生物質が、ベータラクタム、広域ベータラクタム、アミノグリコシド、アンサマイシン、カルバセフェム、カルバペネム、いずれの世代のセファロスポリン、グリコペプチド、リンコサミド、リポペプチド、マクロライド、モノバクタム、ニトロフラン、オキサゾリドノン、ペニシリン、ポリペプチド、キノロン、フルオロキノロン、スルホンアミド、テトラサイクリン、マイコバクテリア抗生物質、クロラムフェニコール、ムピロシンのいずれか1つである請求項32に記載の方法。
- 前記細胞表現型の前記部分が、セフォキシチン、バンコマイシン、テイコプラニン、アンピシリン/スルバクタム、シプロフロキサシン、メロペネム、セフタジジム、セフトリアキソン、ピペラシリン/タゾバクタム、又はゲンタマイシンの少なくとも1つに耐性がある細菌表現型を含む請求項29に記載の方法。
- 前記試薬が抗生物質及び着色剤を含み、前記抗生物質が、前記細胞表現型の前記部分における前記複数のレポーター分子の産生を抑制するように作成され、且つ前記第1の信号が前記着色剤と関連する、請求項29に記載の方法。
- 前記第1の信号が前記反応チャンバー内の前記アッセイ培地の体積に関連する請求項29に記載の方法。
- 前記第2の信号の大きさが所定量を超える複数の形質導入粒子の量に無関係である請求項29に記載の方法。
- 複数の形質導入粒子が、溶解性複製又は溶原性複製のいずれも不能であるように改変される請求項29に記載の方法。
- 前記複数の形質導入粒子がバクテリオファージに由来する請求項29に記載の方法。
- 物質を前記試料に配することを含み、前記物質が前記複数のレポーター分子と反応して第2の信号を生成するように作成される、請求項29に記載の方法。
- 方法であって、
巻いていない材料から構築される採取部分を有する軸を含む綿棒及び前記採取部分から放たれる試料を含む輸送培地を含有する容器を受けること;
前記輸送培地及び前記試料を反応チャンバーに転送すること;
前記反応チャンバー中で前記輸送培地と、標的細胞と関連する、前記標的細胞に複数のレポーター分子を作らせるように作成された核酸分子を含むように改変された複数の形質導入粒子を混ぜること;
前記輸送培地と前記複数の形質導入粒子の前記混合物を少なくとも20℃の温度で約8時間以下の時間維持して、前記標的細胞が前記試料中に存在するとき、複数のレポーター分子を発現させること;並びに
前記複数のレポーター分子の量と関連する信号を受信することを含む方法。 - 前記複数の形質導入粒子が非複製的である請求項41に記載の方法。
- 複数の形質導入粒子が前記複数のレポート分子のレポーター分子を欠いている請求項41に記載の方法。
- 前記複数のレポーター分子のレポーター分子が、細菌ルシフェラーゼ、真核生物ルシフェラーゼ、蛍光タンパク質、比色検出に適した酵素、免疫検出に適したタンパク質、免疫検出に適したペプチド、又はアプタマーとして機能する核酸若しくは酵素活性を呈する核酸のいずれか1つである請求項41に記載の方法。
- 前記維持することが約2時間以下の時間実施される請求項41に記載の方法。
- 前記採取部分が発泡材料から構築される請求項41に記載の方法。
- 前記反応チャンバーが、前記輸送培地と混ざるように作成された凍結乾燥試薬錠剤を含有し、前記凍結乾燥試薬錠剤が抗生物質を含む、請求項41に記載の方法。
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