JP6944999B2 - 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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- JP6944999B2 JP6944999B2 JP2019516521A JP2019516521A JP6944999B2 JP 6944999 B2 JP6944999 B2 JP 6944999B2 JP 2019516521 A JP2019516521 A JP 2019516521A JP 2019516521 A JP2019516521 A JP 2019516521A JP 6944999 B2 JP6944999 B2 JP 6944999B2
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Description
1)サービスプロバイダーに試料収集キットをオンラインで発注する
2)キットを受け取り、生物試料を収集する(通常、唾液である)−この段階で、顧客は、遺伝的特質の結果と併せて分析される質問表への記入を求められる場合がある
3)試料をサービスプロバイダーに郵便で返送する、次に試料は熟練した研究員によって、種々の遺伝的分析の分析評価を用いて処理され、
4)4〜8週間後に、顧客に、顧客のDNAにおける異なる変異を羅列した包括的分析報告が電子または郵便で送達される。場合によっては、サービスプロバイダーは、依頼人の遺伝的プロファイルに従って設計または選択された特注製品の製品推奨または提案を行ってもよい。
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。
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。
・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。
に記載されたような挿入色素を利用してよい。別の手法はプローブベースの手法、例えばhttps://www.thermofisher.com/uk/en/home/life−science/pcr/real−time−pcr/real−time−pcr−assays/snp−genotyping−taqman−assays.htmlを含む。
結合ステーションを介して、スマートフォンはベースユニットと通信し、ベースユニットに、SNP遺伝子型決定手順の種々のステップを実行するために、第1と第2のモータ、ヒーター等を作動させるようにする。手順中の適切な時点で、スマートフォンに信号が送信されて、スマートフォンに自身のカメラを作動させる。いくつかの場合では、検査終了時に単一の画像のみがキャプチャされるが、手順を通じて多数の画像をキャプチャすることによって、検査結果への特異度が増強され得る。
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 (10)
- DNAまたはRNAを含む生物材料の試料を調製してPCRを実行するためのシステムであって、
テストカートリッジと、
ベースユニットと、を備え、
前記テストカートリッジは、
フロースルーチャンバを画定する第1のハウジングと、
前記第1のハウジングが少なくとも部分的に内部に配置される中央空間を画定する第2のハウジングとを有し、
前記第1のハウジングは前記第2のハウジングに対して回転可能であり、前記第2のハウジングは複数の周方向に離間したチャンバを画定し、前記チャンバのうち1つは試料を受け入れるための取入口を有し、前記チャンバのうち少なくとも1つは液体試薬を保持しており、前記チャンバのうち少なくとも1つは前記第2のハウジングに差し込まれるまたは前記第2のハウジングと一体に形成される分析チャンバであり、前記分析チャンバは前記PCRを実行するためのものであり、前記第2のハウジングのチャンバはそれぞれ前記中央空間への開口部を有し、
前記第1のハウジングは、前記中央空間への1つ以上の開口部を有し、その結果、その開口部または各開口部が、前記第1のハウジングと前記第2のハウジングの相対回転によって、前記第2のハウジングのチャンバへの開口部のうち1つと選択的に整列されることができ、
前記ベースユニットは、
前記第1のハウジングが前記ベースユニットに対して回転可能でありながら、前記第2のハウジングが前記ベースユニットに対して回転できないように、前記テストカートリッジを前記ベースユニットに結合するための構造を画定する、またはそのような構造が接続されたハウジングと、
前記分析チャンバを加熱するために、前記分析チャンバの下に配置されているヒーターモジュールと、
前記第1のハウジングの前記第2のハウジング内での回転を引き起こさせるために前記第1のハウジングと係合する第1のドライバと、
前記第1のハウジングの前記フロースルーチャンバに正圧および負圧をかける第2のドライバと、
前記第1と第2のドライバを作動させて、前記チャンバ間での液体の所望のシーケンスでの変位を引き起こし、前記分析チャンバへの、調製された試料の配給をもたらしてそれによって前記分析チャンバによる分析結果の提供を促進するコントローラと、
前記分析チャンバにおいて生成された視覚的に検出可能な分析結果の上から撮像を可能にするために前記分析チャンバの上を摺動可能な光学素子モジュールと、を有し、
前記分析チャンバは前記分析チャンバの周囲に固定された透明プラスチックシートにより覆われており、前記分析チャンバは、前記透明プラスチックシートを通して前記テストカートリッジの上から見える、視覚的に検出可能な分析結果を提供するように構成され、
前記分析チャンバは、PCRが実行可能であり、前記視覚的な結果が各ウェルにおける1つ以上のバイオマーカーから生成される、ウェルのアレイを有しており、前記ウェルの基部は、空気を通すが液体透過性ではない材料によって形成され、前記透明プラスチックシートは、分析のために調製された試料を受け入れるための前記ウェルの上の空間を画定するためにウェルの上にあり、
前記ベースユニットは、さらに、前記分析チャンバの底を上方に偏向させるために前記ヒーターモジュールと係合するリニアアクチュエータを有し、前記分析チャンバの上部を前記垂直方向に運動できない前記光学素子モジュールの底に押し付け、それによって前記ウェルの上の液体に圧力を及ぼして、満足な充填レベルを達成するために前記液体を前記ウェルに押し入れる、システム。 - 前記第2のハウジングはさらに、排液を抜くまたは受けるための少なくとも1つの空チャンバを画定する、請求項1に記載のシステム。
- 試料を受けるための前記取入口はスワブの試料ヘッドを受けるように構成されている、請求項1または2に記載のシステム。
- 前記システムは一塩基多型遺伝子型決定を実行するように構成されてよく、前記テストカートリッジのチャンバは、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3のいずれか一項に記載のシステム。
- 前記分析チャンバはDNA増幅を実行するように構成され、増幅を示すインターフェースを有する、請求項4に記載のシステム。
- 前記システムとスマートフォンとの結合を促進するための構造を備え、その結果、前記分析チャンバの分析結果が前記スマートフォンのカメラによって撮像され得る、請求項1乃至5のいずれか一項に記載のシステム。
- 前記第1のハウジングは、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6のいずれか一項に記載のシステム。
- 前記多孔膜はシリカフリットである、請求項7に記載のシステム。
- 前記第1のハウジングは、中央空間への前記開口部のうち2つ以上を備え、前記開口部のうち少なくとも1つは妨げられず、もう1つの開口部は前記多孔膜によって妨げられる、請求項7または8に記載のシステム。
- 前記妨げられた開口部と妨げられない開口部のうち一方が、前記第2のハウジングのチャンバへの開口部のうち1つと整列すると、妨げられた開口部と妨げられない開口部のうち他方はそのように整列しないように構成される請求項9に記載のシステム。
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