JP7451779B2 - 血液分析装置および血液分析方法 - Google Patents
血液分析装置および血液分析方法 Download PDFInfo
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Description
実施形態1の血液分析装置10は、血液検体と試薬とから調製された測定試料を測定して、マラリア原虫に感染した赤血球を含む集団を特定し、血液検体がマラリアに感染しているか否かを判定する。
発明者らが、図5(b)に示すマラリア感染赤血球に対応する領域203に、細胞外小胞の一種であるアポトーシス小体と、核酸の封入体の一種であるハウウェルジョリー小体を含む赤血球とが分布することを見いだした経緯について説明する。核酸の封入体を含む赤血球とは、細胞の内部で核酸が凝集して残った核酸の封入体を含む赤血球である。なお、網赤血球においては、内部に存在するRNAは網赤血球内で分散しており、核酸は凝集体としては存在しない。したがって、網赤血球は、核酸の封入体を含む赤血球とは異なるものである。
・培地:10% FBS添加 RPMI1640培地
・細胞溶液:1×106/μL程度まで培養したJurkat細胞を培地で10倍希釈し、24時間培養
(アポトーシス誘導)
・誘導剤:Staurosporine 1μM
・Stock solution:250μg/500μL DMSO(1mMStaurosporine)
・1mM Staurosporineを培地で10倍希釈した100μM Staurosporine溶液を作成し、培養液に対して1/100量添加
実施形態2では、判定処理において、マラリアの有無だけでなく、核酸の封入体を含む赤血球、および細胞外小胞の少なくとも一方の有無についても判定される。実施形態2および後述する実施形態3において「核酸の封入体を含む赤血球、および細胞外小胞の少なくとも一方の有無を判定する」とは、核酸の封入体を含む赤血球、または細胞外小胞の有無を判定すること、すなわち核酸の封入体を含む赤血球、および細胞外小胞のうち少なくとも一方があるか否かを判定することを示す。実施形態2の判定処理は、図6に示した実施形態1の判定処理と比較して、判定内容が異なっているものの、図16に示すように、略同様に構成されている。実施形態2のその他の構成は、実施形態1と同様である。
実施形態3では、判定処理において、核酸の封入体を含む赤血球、および細胞外小胞の少なくとも一方の有無について判定される。上述したように、スキャッタグラム200において、マラリア感染赤血球に対応する領域203には、核酸の封入体を含む赤血球、または細胞外小胞が分布する場合がある。実施形態3では、図18に示すように、スキャッタグラム400において、核酸の封入体を含む赤血球、および細胞外小胞に対応する領域403が設けられている。また、実施形態3の判定処理は、図19に示すように構成される。実施形態3のその他の構成は、実施形態1と同様である。
30 試料調製部
51 制御部
53 出力部
53a 表示部
101 フローセル
102 光源
103、105 受光部
400 スキャッタグラム
403 領域
500 度数分布図
G6 粒子群
Claims (20)
- 血液検体と核酸を染色する蛍光色素とを混合して測定試料を調製し、調製された前記測定試料に光を照射して前記測定試料中の粒子から生じる蛍光強度および散乱光強度を含む測定データを生成する測定部と、
前記測定データに含まれる蛍光強度および散乱光強度に基づいて特徴空間に粒子をプロットし、前記特徴空間においてマラリア感染赤血球が現れる領域にプロットされる粒子を計数する制御部と、を備え、
前記制御部は、第1の量の前記測定試料を測定して得られた第1の測定データに基づいて前記マラリア感染赤血球が現れる領域にプロットされる粒子の数が閾値未満である場合、前記第1の量より多い第2の量の前記測定試料を測定するよう前記測定部を制御する、血液分析装置。 - 前記制御部は、前記測定部が前記第2の量の測定試料を測定して得られた第2の測定データに基づいて、マラリア原虫の感染に関する判定結果を取得する、請求項1に記載の血液分析装置。
- 前記制御部は、前記第1の測定データに基づいて前記マラリア感染赤血球が現れる領域にプロットされる粒子の数が閾値以上である場合、前記第1の測定データに基づいてマラリア原虫の感染に関する判定結果を取得する、請求項1に記載の血液分析装置。
- 前記制御部は、前記特徴空間においてシングルリングフォーム赤血球が現れる領域にプロットされる粒子の分布のばらつきを示す値を取得し、その値に基づいてマラリア原虫の感染に関する判定結果を取得する、請求項1に記載の血液分析装置。
- 前記制御部は、前記ばらつきを示す値と、マラリア感染赤血球が現れる領域に含まれる粒子数とに基づいて、マラリア原虫の感染の有無に関する判定結果を取得する、請求項4に記載の血液分析装置。
- 前記ばらつきを示す値は、シングルリングフォーム赤血球が現れる領域にプロットされる粒子の蛍光強度に対する度数分布のばらつきを示す値である、請求項5に記載の血液分析装置。
- 前記ばらつきを示す値は、変動係数、標準偏差または度数分布図の半値幅である、請求項5に記載の血液分析装置。
- 前記試料調製部は、赤血球を収縮させる希釈液を混合して前記測定試料を調製する、請求項1に記載の血液分析装置。
- 前記光源は、青紫色波長帯域の光を出射する、請求項1に記載の血液分析装置。
- 前記蛍光強度および前記散乱光強度に基づくスキャッタグラムを表示する表示部をさらに備える、請求項1に記載の血液分析装置。
- 血液検体と核酸を染色する蛍光色素とを混合して測定試料を調製し、
調製された前記測定試料のうち第1の量の測定試料をフローセルに流し、前記フローセルを流れる前記測定試料に対して光を照射し、
前記測定試料中の粒子から生じる蛍光強度および散乱光強度を測定し、
蛍光強度および散乱光強度に基づいて特徴空間に粒子をプロットし、
前記特徴空間においてマラリア感染赤血球が現れる領域にプロットされる粒子の数が閾値未満である場合、前記第1の量より多い第2の量の測定試料を前記フローセルに流し、第2の量の測定試料中の粒子から得られる蛍光強度および散乱光強度に基づいて、前記血液検体中のマラリア感染赤血球の有無を判定する、血液分析方法。 - 前記第2の量の測定試料を測定して得られた、前記マラリア感染赤血球が現れる領域にプロットされる粒子の数に基づいて、前記血液検体中のマラリア感染赤血球の有無を判定する、請求項11に記載の血液分析方法。
- 第1の量の測定試料を測定したときの前記マラリア感染赤血球が現れる領域にプロットされる粒子の数が閾値以上である場合、前記第1の量の測定試料中の粒子から得られる蛍光強度および散乱光強度に基づいてマラリア原虫の感染に関する判定結果を取得する、請求項11に記載の血液分析方法。
- 前記特徴空間においてシングルリングフォーム赤血球が現れる領域にプロットされる粒子の分布のばらつきを示す値を取得し、その値に基づいて前記血液検体中のマラリア感染赤血球の有無を判定する、請求項11に記載の血液分析方法。
- 前記ばらつきを示す値と、マラリア感染赤血球が現れる領域に含まれる粒子数とに基づいて、前記血液検体中のマラリア感染赤血球の有無を判定する、請求項14に記載の血液分析方法。
- 前記ばらつきを示す値は、シングルリングフォーム赤血球が現れる領域にプロットされる粒子の蛍光強度に対する度数分布のばらつきを示す値である、請求項15に記載の血液分析方法。
- 前記ばらつきを示す値は、変動係数、標準偏差または度数分布図の半値幅である、請求項15に記載の血液分析方法。
- 前記測定試料の調製は、血液検体にさらに赤血球を収縮させる希釈液を混合することを含む、請求項11に記載の血液分析方法。
- 前記測定試料に照射される光は、青紫色波長帯域の光である、請求項11に記載の血液分析方法。
- 前記蛍光強度および前記散乱光強度に基づくスキャッタグラムを表示することをさらに含む、請求項11に記載の血液分析方法。
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