JP2022001873A - 生物試料の分析方法および装置 - Google Patents
生物試料の分析方法および装置 Download PDFInfo
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Abstract
Description
デバイスは、遺伝子発現および単純な後成的検査などの他の分析評価にも用いられてよい。本発明は、個人的な遺伝およびその他の生物データをユーザデバイスに安全に保存して使用する方法および装置にも関する。
1)誰が生物試料を分析するか、また、分析がどのようにどこで行われるか
2)遺伝子情報がどのように且つどこに保存され、または安全に破棄されるか
3)顧客の個人情報が顧客の遺伝子プロファイルとどのようにリンクされるか
4)顧客の遺伝的情報に誰がアクセスできるか
5)多くの場合、遺伝的分析報告を出す前に、顧客は、自身の病歴、生活習慣等に関する質問表に記入するように求められるが、そのような情報を共有することで別次元の懸念が増える。
Interface Science,181,635−644(1996)に記載されている。別法として、DNAはシリカコーティングされた磁性粒子上に捕捉されて、磁界の印加によって側壁に収束されることによって回転チャンバから離れることを防止されてよい。DNAを結合するために磁性粒子と磁界を使用する一例のシステムは、米国マサチューセッツ州ウォルサムのサーモフィッシャー(Thermo Fisher)社から入手可能である。
チャンバ2/溶解チャンバ(5b):溶解バッファを保持する。SNP遺伝子型決定において、溶解バッファは、細胞膜を壊して被検体のDNAを解放するために配設されている。
チャンバ3/洗浄チャンバ(5c):洗浄液試薬を保持する。これは、例えば、エタノールであってよい。
チャンバ4/溶出チャンバ(5d):このチャンバは、脱イオン水または10mMTrispH8.5などの固相からDNAを除去するための溶出試薬を保持する。
チャンバ5/排出・通気チャンバ(5e):このチャンバは使用前に空であり、下部ユニット(排出サンプ)と液体連通している。
チャンバ6/分析チャンバ(5f):このチャンバは、増幅とシーケンシングを実行するように構成されたチップモジュールを保持する。例として、チップは、英国ロンドンのDNAeによって開発されたシステムのようなISFETベースの検出システムを使用してよい。チップモジュールは、充填された試薬チャンバ、マイクロ流体、1つのまたは複数のヒーターおよび制御エレクトロニクスを含む種々の構成要素を備えている。
・液体を上部ハウジングのチャンバから受け、所望の期間にわたり回転ハウジング内に保持する(それによって反応ステップを分離する)ことが可能である。回転チャンバは、上部ハウジングのチャンバのいずれの開口部も回転チャンバへの開口部に整列しないように回転され得る。
・複数の試薬が、プロトコルの特定の時点で混合される(試薬が長期間にわたり混ざって保管されることが望ましくない場合、または希釈が必要な場合)。これは、回転チャンバ内または上部ハウジングのチャンバのうち1つ以上において実行され得る。
・回転チャンバ内に固相(シリカフリット)を配設することは、チャンバが単に単純な「試薬転移」要素ではなく、寧ろプロセスにおいて能動的な機能参与因子であるということを意味する。
・ピストンの回転チャンバとの相互作用は、例えば、液体を回転チャンバ、上部ハウジングのチャンバ間で圧送して再び戻すことによって、混合動作を組み込むことを提供する。
・完全に展開していない場合のピストンの相互作用は、回転チャンバと上部ハウジングのチャンバとの間の容積、または上部ハウジングと回転チャンバとの間の容積の100%未満を分配するために用いられてよい。
・回転チャンバには2以上の開口部が配設されてよい。実際、図面に示された例は、2つのそのような開口部を含む。これらは、上部ハウジングのチャンバ内の開口部と選択的に整列するために周方向に離間している。
主ねじ31がその中で運動する円筒形内部空間には流出しないということが明らかとなろう。したがって、上向きに延出する円筒形壁17は「堰」として働き、内部空間の汚染、また重要なことには主ねじとキャプチャヘッドの汚染を防止する。これにより、ベースユニットの汚染がないことが保証され、さらなるテストカートリッジでのベースユニットの再利用を可能にする。
・インプット−試料ヘッドが試料チャンバに導入される。ヘッドが折り取られて試料チャンバを封止する。試料処理システムはベースユニット上に配置されている。回転チャンバは換気されている。図11a。
・溶解−回転チャンバが回転されて、溶解試薬を保持するチャンバに接続される。溶解試薬は試薬チャンバから回転チャンバに転移される。回転チャンバは回転して試料チャンバに接続する。溶解試薬が回転チャンバから試料チャンバに転移される。試料チャンバ内の試薬は上昇した温度まで加熱されて細胞内部からDNAを取り出すことを補助する。DNAを保持した溶解試薬が試料チャンバから回転チャンバに転移される。回転チャンバが回転して排出チャンバに接続する。DNAを保持した溶解試薬はDNA捕集材料に接触する。上清が回転チャンバから排出チャンバに転移される。図11b。
・洗浄−回転チャンバが回転されて洗浄試薬を保持するチャンバに接続される。洗浄試薬は試薬チャンバから回転チャンバに転移される。回転チャンバは回転して排出チャンバに接続する。洗浄試薬がDNA捕集材料に接触する。上清が回転チャンバから排出チャンバに転移される。図11c。
・溶出−回転チャンバが回転されて溶出試薬を保持するチャンバに接続される。溶出試薬が試薬チャンバから回転チャンバに転移される。回転チャンバは、フリットを保持する開口部を分析チャンバに接続する。溶出試薬がDNA捕集材料(多孔フリット)を通過し、溶液にDNAを放出する。DNAを含む溶出試薬が回転チャンバから分析チャンバに転移される。図11d。
・増幅−DNAを含む溶出試薬が上昇した温度に加熱されて内部のDNAを増幅する。増幅は、等温または温度サイクリング(PCRなど)のいくつかの既知の異なる方法のうち1つを用いる可能性がある。複数のプライマーおよびプローブセットが、指定されたDNA増幅プロトコルで動作するように設計されたチップモジュールの各アリコートに存在してよい。最適な動作(例えば、効率と速度に関して)のため、プライマーは、短いアンプリコンを生成するような方式に設計され得る。図11e。
2 ベースユニット
3 テストカートリッジ
4 ボタン
5 上部マルチチャンバユニット
5a−f チャンバ
6 下部ユニット
7 回転チャンバ
8 半径方向に延出する壁
9 内部空間
10 閉じた上面
11 円筒形壁
12 半径方向に延出する穴
13 円形開口部
14 円筒形部材
15 上部壁
16 円形開口部
17 円筒形壁
18 内向きに突出するリップ
19 ベベル形状歯車
20 プランジャヘッド
21 捕捉形状
22 構成要素(チップモジュール)
23 上部ユニットの開いたセグメント
24 外部ハウジング
25 第1の回転モータ
26 ベベル形状ピニオン
27 コントローラ
28 第2の回転モータ
29 第1の歯車
30 第2の歯車
31 主ねじ
32 キャプチャヘッド
33 半径方向に延出する 開口部
34 口内スワブ
35 口内スワブ捕集端部
36 口内スワブハンドル
37 アクセスポート
50 ベースユニット(第2の実施形態)
51 結合ステーション
52 スマートフォン
53 テストカートリッジ
54 カートリッジ受け入れポート
55 スマートフォン
56 器具
57 結合コンパートメント
58 カメラ
59 コントローラ
60 器具メモリ
61 スマートフォンメモリ
62 製品データベース
63 スマートフォンプロセッサ
64 シリカフリット
65 ベースユニット
66 光学素子
67 PCB
68 回転ステッパモータ
69 注射器
70 テストカートリッジ
71 分析チャンバ
72 ペルチェモジュール
73 スマートフォン
74 回転チャンバ
75 マルチチャンバユニット
76 上部壁構成要素
77 ロックナット
78 ばね
78a ばねポケット
79 フロースルーチャンバへの開口部
80 Oリング
81 液体
82 円筒形部材
83 二方弁
84 ウェルのアレイ
85 ウェル基部
86 リニアアクチュエータ
87 中間構成要素
88 カム
89 貫通アーム
90 水槽
91 取入口開口部
92 主開口部
93 排出開口部
94 ウェル射出開口部
95 凍結乾燥された粒子
96 マルチチャンバユニットの開き口
97 マルチチャンバユニットに配設された経路
Claims (14)
- 生物材料の試料を分析するためのテストカートリッジであって、
複数のチャンバと1つの中央空間を画定する外部ハウジングと、
前記チャンバと共に前記中央空間の周りに周方向に配置されているモジュールと、
前記中央空間に配置されてフロースルーチャンバを画定する内部ハウジングとを備え、
前記内部ハウジングは外部ハウジング内で回転可能であり、
前記外部ハウジングは前記中央空間と前記複数のチャンバの各々との間の複数の開口部と、前記モジュールと連通するさらなる1つの開口部を画定し、
前記モジュールは、
前記生物材料の試料または前記生物材料の試料由来成分を含有した液体を受け入れるためのさらなる開口部と液体連通する取入口と、前記ベース部材を通って延出し分析が実行されるウェルのアレイとを画定するベース部材と、
空気と液体の両方に不透過であり、前記ベース部材の上に固定されて前記ウェルのアレイの上に及び前記ウェルが開口する、前記取入口と液体連通する第1の空間を定義する、透明な可撓性の膜と、
前記ベース部材に溶接されて前記ウェルのアレイの下に第2の空間を定義する、エラストマーのさらなる空気および液体不透過膜と、
前記ウェルのアレイの下に固定されて前記第2の空間から前記ウェルを分離する液体不透過および空気透過膜と、
前記ウェルに利用可能となるように前記液体不透過および空気透過膜に固定された1つ以上のバイオマーカーとを有する、テストカートリッジ。 - 前記外部ハウジングは、前記内部ハウジングの回転軸と整列した第1の略円錐台面を画定し、前記内部ハウジングは前記第1の略円錐台面に当接する第2の略円錐台面を画定し、前記開口部は前記第1の略円錐台面内に配設され、前記フロースルーチャンバへの1つの開口部が前記第2の略円錐台面内に画定され、前記テストカートリッジは、前記フロースルーチャンバへの開口部の、前記外部ハウジング内に画定された前記開口部のいずれか1つへの整列を可能にするように構成される、請求項1に記載のテストカートリッジ。
- 前記内部ハウジングの前記第2の略円錐台面に対して、前記外部ハウジング内に画定された前記開口部を封止するための1つまたは複数のシールを備えている、請求項2に記載のテストカートリッジ。
- 前記第1の略円錐台面と前記第2の略円錐台面を互いに対して圧迫するための付勢部材をさらに備えている、請求項3に記載のテストカートリッジ。
- 前記外部ハウジングのチャンバを覆う1つ以上の他の可撓性の膜を備え、
さらに、前記1つ以上の他の膜に貫通してそれらを周囲環境に通気するために外部ハウジングに対して軸方向に可動な構成要素を備えている、請求項1乃至4のいずれか一項に記載のテストカートリッジ。 - 前記構成要素は、前記内部ハウジングが回転すると前記外部ハウジングに相対して軸方向に可動となるように構成されている、請求項5に記載のテストカートリッジ。
- 前記ウェルはそれぞれ、各ウェルが底部よりも頂部において幅狭となるように、1つ以上の傾斜した側壁によって画定されている、請求項1乃至6のいずれか一項に記載のテストカートリッジ。
- 前記チャンバは、溶解バッファを保持した少なくとも1つのチャンバと、洗浄液試薬を保持するチャンバと、溶出試薬を含むチャンバを備えている、請求項1乃至7のいずれか一項に記載のテストカートリッジ。
- 前記チャンバは、排液を抜くまたは受けるための少なくとも1つの空チャンバを有する、請求項1乃至8のいずれか一項に記載のテストカートリッジ。
- 前記テストカートリッジは、前記生物試料に含まれるDNAまたはRNAを検査するためのものである、請求項1乃至9のいずれか一項に記載のテストカートリッジ。
- 前記複数のチャンバのいくつかは、スワブの試料ヘッドを受けるように構成されている取入口を備えている、請求項1乃至10のいずれか一項に記載のテストカートリッジ。
- 前記モジュールはさらに、前記アレイに固定されて前記ウェルを前記第1の空間から分離する空気および液体透過膜を備えている、請求項1乃至11のいずれか一項に記載のテストカートリッジ。
- 前記内部ハウジングは、前記フロースルーチャンバ内の、または前記フロースルーチャンバと液体連通する多孔膜または他の構造を備え、前記多孔膜はDNA材料を保持するように構成される、請求項1乃至12のいずれか一項に記載のテストカートリッジ。
- 前記内部ハウジングは、空気が圧力下で前記内部ハウジングに入る方向と出る方向に通ることを可能にするが、飛沫が前記内部ハウジングの上部に移動することは防止するように構成された二方弁を備えている、請求項1乃至13のいずれか一項に記載のテストカートリッジ。
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