JP7162628B6 - 1種類以上の蛍光シグナルをリアルタイムで検出するポリメラーゼ連鎖反応装置 - Google Patents
1種類以上の蛍光シグナルをリアルタイムで検出するポリメラーゼ連鎖反応装置 Download PDFInfo
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Description
Claims (24)
- 試薬容器における1種類以上の蛍光シグナルをリアルタイムで検出し、且つ定量するポリメラーゼ連鎖反応(PCR)装置であって、前記試薬容器は、反応物、及び1種類以上の蛍光プローブ又は蛍光染料を含む反応試薬を含み、前記PCR装置は、
前記試薬容器を収容する試薬容器収容機構と、
前記試薬容器収容機構の一方側に位置しており、前記試薬容器に入射し、且つ蛍光を生成するように前記1種類以上の蛍光プローブ又は前記蛍光染料を励起するための光線を発する光源と、
前記試薬容器収容機構の一方側に位置しており、1種類以上の前記蛍光の蛍光シグナルを検出するように設けられる分光計と、
前記1種類以上の蛍光シグナルを定量するためのアルゴリズムがプリロードされており、前記試薬容器収容機構の昇温と降温の周期、及び前記光源のオン/オフの時点を制御する処理部と、を含み、
(イ)前記試薬容器収容機構は、第1凹面を有する第1基板、第2凹面を有する第2基板、第1導電性部材、及び第2導電性部材により構成され、前記第1導電性部材及び前記第2導電性部材は、前記第1基板の2つの異なる端部に設けられ、且つ前記第1基板と前記第2基板により挟持されており、前記第1凹面と前記第2凹面とは、前記試薬容器が収容される空間を形成するように対向しており、前記第1基板及び前記第2基板は耐熱性物質により構成され、前記耐熱性物質は金属又は非金属であり、或いは、
(ロ)前記試薬容器収容機構は、第3基板、第4基板及び絶縁シートにより構成され、前記第3基板及び前記第4基板は、耐熱性物質により構成され、所定の形状に湾曲され、前記耐熱性物質は金属であり、前記第3基板と前記第4基板とは鏡面対称になるように組み合わせられ、前記絶縁シートは、前記第3基板と前記第4基板との間に配置されている、PCR装置。 - 前記試薬容器収容機構が通電により加熱を開始し、前記試薬容器が昇温し、
前記処理部が前記試薬容器収容機構の温度の第1温度範囲と第2温度範囲との間のサイクルを制御し、前記反応物と前記反応試薬とがPCRを行い、
前記分光計により検出された前記1種類以上の蛍光シグナルが前記処理部に伝送されて解析される、請求項1に記載のPCR装置。 - 放熱部と、
前記試薬容器収容機構の一方側に位置しており、前記試薬容器収容機構の温度を測定する温度センサと、をさらに含み、
前記試薬容器収容機構の温度が前記第1温度範囲を超えたと前記温度センサにより検出された場合、前記温度センサが前記処理部にフィードバックし、降温を行うように前記処理部が前記放熱部に第1信号を送信し、
前記試薬容器収容機構の温度が前記第2温度範囲よりも低いと前記温度センサにより検出された場合、前記温度センサが前記処理部に情報をフィードバックし、通電により加熱を開始するように前記処理部が前記試薬容器収容機構に第2信号を送信する、請求項2に記載のPCR装置。 - 前記試薬容器収容機構の温度は、リアルタイムで検出される、請求項3に記載のPCR装置。
- 前記放熱部は、1つ以上のファン、1つ以上の熱電冷却器及びその組み合わせにより構成された群から選択されたものである、請求項3に記載のPCR装置。
- 前記試薬容器収容機構と前記光源との間に位置しており、前記光線を前記試薬容器に導き、且つ蛍光を生成するように前記1種類以上の蛍光プローブ又は前記蛍光染料を励起する第1光学素子、をさらに含み、
前記第1光学素子及び前記光源は、前記試薬容器収容機構の下方に順次配置されている、請求項1に記載のPCR装置。 - 前記試薬容器収容機構と前記分光計との間に位置しており、蛍光を収集し、且つ前記蛍光を前記分光計に導く第2光学素子、をさらに含み、
前記第2光学素子及び前記分光計は、同一の水平面に位置する、請求項6に記載のPCR装置。 - 前記第1光学素子及び前記第2光学素子のそれぞれは、両凸レンズ、平凸レンズ、二重レンズ、非球面レンズ、色消しレンズ、収差消しレンズ、フレネルレンズ(Fresnel lens)、平凹レンズ、両凹レンズ、正/負メニスカスレンズ、アキシコン、屈折率分布型レンズ、マイクロレンズアレイ、レンチキュラーレンズ、回折光学素子、導波路素子(waveguide elements)、ホログラフィック光学素子、鏡、繊維及びプリズムにより構成された群から選択されたものである、請求項7に記載のPCR装置。
- 前記第1導電性部材及び前記第2導電性部材は、金属シート又はプリント回路基板(printed circuit boards:PCBs)である、請求項1に記載のPCR装置。
- 前記第1導電性部材及び前記第2導電性部材が金属シートである場合、前記第1基板及び前記第2基板のそれぞれの表層は導電性フィルムをさらに含み、前記導電性フィルムは、酸化スズ、酸化インジウム、酸化亜鉛、酸化インジウムスズ、クロム、チタン、タンタル又は銅により構成された群から選択された材質により形成される、請求項9に記載のPCR装置。
- 前記第1導電性部材及び前記第2導電性部材がプリント回路基板(printed circuit boards:PCBs)である場合、前記プリント回路基板は1つ以上の電熱素子をさらに含み、前記電熱素子は抵抗器又はPCB回路配線である、請求項9に記載のPCR装置。
- 前記第1基板と前記第2基板とは、前記第1凹面と前記第2凹面とが水平面に略垂直な方向に結合面に沿って互いに結合して前記試薬容器収容機構を形成するように、互いに平行に配置されている、請求項1に記載のPCR装置。
- 前記第1基板は、蛍光シグナル出口窓を有し、
前記第1基板は、前記第2基板と前記分光計との間に位置する、請求項1に記載のPCR装置。 - 前記第1基板及び前記第2基板は、1つ以上の放熱孔を有し、
前記耐熱性物質は、金属であり、アルミニウム、銅、アルミニウム銅合金及び他の耐熱性金属により構成された群から選択されたものである、請求項1に記載のPCR装置。 - 前記耐熱性物質は、金属であり、
前記耐熱性物質の表層は、耐熱絶縁性物質を含み、
前記耐熱絶縁性物質は、酸化アルミニウム、ポリテトラフルオロエチレン(polytetrafluoroethylene)及びポリイミド(polyimide)により構成された群から選択されたものである、請求項14に記載のPCR装置。 - 前記耐熱性物質は、非金属であり、ガラス、プラスチック及びセラミックにより構成された群から選択されたものである、請求項1に記載のPCR装置。
- 前記アルゴリズムは、最小二乗法を用いるアルゴリズムである、請求項1に記載のPCR装置。
- 前記光源は、LED(light emitting diode)、レーザ光源、異なる波長を有する複数のLED、異なる波長を有する複数のレーザ光源及びその組み合わせにより構成された群から選択されたものである、請求項1に記載のPCR装置。
- 前記処理部は、前記光源が所定の時間間隔でオン又はオフになるように前記光源を制御し、
前記分光計は、前記光源のオン期間に同期して前記1種類以上の蛍光シグナルを検出する、請求項1に記載のPCR装置。 - 前記分光計の検出範囲は、340nm~850nmである、請求項1に記載の装置。
- 前記試薬容器収容機構と前記分光計との間に位置する光フィルタ、をさらに含み、
前記光フィルタは、前記1種類以上の蛍光シグナルの範囲を超えた波長を有する光を除去する、請求項1に記載のPCR装置。 - 340nm~850nmの波長を有する前記1種類以上の蛍光プローブ又は前記蛍光染料は、FAM、VIC、HEX、ROX、CY3、CY5、CY5.5、JOE、TET、SYBR、Texas Red、TAMRA、NED、Quasar705、Alexa488、Alexa546、Alexa594、Alexa633、Alexa643又はAlexa680である、請求項1に記載のPCR装置。
- 前記第1温度範囲は85℃~130℃であり、
前記第2温度範囲は50℃~75℃である、請求項2に記載のPCR装置。 - 前記処理部は、前記第1温度範囲及び前記第2温度範囲とは異なる第3温度範囲を有するように構成され、
前記処理部は、前記試薬容器収容機構の温度を前記第1温度範囲、前記第2温度範囲及び前記第3温度範囲で繰り返し昇温/降温させるように制御する、請求項2に記載のPCR装置。
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