JP5230648B2 - 自動血液分析装置による全血試料中の有核赤血球及び白血球細胞の測定方法 - Google Patents
自動血液分析装置による全血試料中の有核赤血球及び白血球細胞の測定方法 Download PDFInfo
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Description
本発明の方法は、特別な溶解試薬と、有核赤血球濃度と白血球濃度の両方を測定する光散乱多角度偏光解消(multi−angle depolarizing)フローサイトメーターとの組合せを使用することができる。本方法は、有核赤血球集団を白血球集団から分離すること、及び例えば、血小板凝集塊、脂質、粒子の形の溶解した赤血球などの他の干渉粒子を同定することを含む。
(a)溶解された全血試料を用意する段階、
(b)溶解試料を光散乱多角度偏光解消フローサイトメーターに導入する段階、
(c)偏光解消干渉(depolarizing interference)、例えば、脂肪滴及び他の測定される粒子を除去する段階、
(d)偏光解消干渉の非存在下で、有核赤血球及びノイズを白血球から識別する段階、
(e)偏光解消干渉及び白血球の非存在下で、有核赤血球をノイズから識別する段階、及び
(f)存在し得る血小板凝集塊を識別する段階
を含む。
本明細書では「形態学的評価」という表現は、細胞の形状の評価を意味する。「白血球細胞」という用語は、白血球を意味する。赤血球とは異なり、白血球は多数の異なるタイプで存在する。白血球細胞の例としては、顆粒球、好中球、好酸球、好塩基球、リンパ球及び単球が挙げられる。「参照方法」という表現は、試験方法を比較する対象である従来技術の方法を意味する。「鎌状赤血球」という用語は、鎌のような形状の赤血球を意味する。鎌状赤血球は、典型的には、溶解試薬に対して耐性がある。「地中海貧血症」という用語は、骨髄が十分な赤血球を形成できず、赤血球寿命も短縮する、遺伝性血液疾患に関係する。「リンパ球」という用語は、リンパ節、ひ臓及び他のリンパ組織中で成熟し、血液に入り、全身を循環する白血球を意味する。「有核赤血球」という表現は、依然として核を含む未成熟赤血球を意味する。本明細書では「ノイズ」という用語は、粒子の形の溶解した赤血球、細胞片、血小板凝集塊などの物質を含むが、これらだけに限定されない。
(1)0°及び10°検出器によって収集されるデータを用いて除外される、粒子の形の溶解した赤血球。
(2)90°偏光解消及び0°検出器によって収集されるデータを用いて除外される、脂質。
(3)90°検出器によって収集されるデータを用いて除外される、血小板凝集塊。
(1)初回の報告可能な白血球の増加(失効の減少)、
(2)血小板凝集塊の高感度計数、
(3)脂肪滴の高感度計数
が得られる。
(1)0°光散乱:レーザー光線に対して約1°から約3°で散乱する光。
(2)10°光散乱:レーザー光線に対して約3°から約10°で散乱する光。
(3)90°光散乱:レーザー光線に直交して散乱する光。
(4)90°偏光解消光散乱:白血球と相互作用することによってもはや垂直に偏光しない、レーザー光線に直交して散乱する光。
Claims (5)
- (a)溶解された全血試料を用意する段階、
(b)溶解試料を多重散乱偏光解消フローサイトメーターにかける段階、
(c)フローサイトメーターの90°偏光解消散乱及び0°散乱検出器によって収集されたデータを用いて偏光解消干渉を除去する段階、
(d)偏光解消干渉の非存在下で、有核赤血球及びノイズを白血球から識別する段階、
(e)偏光解消干渉及び白血球の非存在下で、有核赤血球をノイズから識別する段階、及び
(f)存在し得る血小板凝集塊を白血球から識別する段階、並びに
(g)溶解された全血試料中の有核赤血球数及び全白血球数を計算する段階
を含む、白血球分類分析を要求することなく、同じ血液試料からの有核赤血球及び全白血球の計数のための方法。 - ノイズが、0°散乱及び10°散乱検出器によって収集されたデータを用いて除去される、請求項1に記載の方法。
- 偏光解消干渉が、脂質からの干渉を含む、請求項1に記載の方法。
- 血小板凝集塊が、90°散乱検出器によって収集されたデータを用いて除去される、請求項1に記載の方法。
- 溶解された全血試料が、希釈剤及び溶解試薬によって溶解されており、溶解試薬と希釈剤の比が1:3から1:8の範囲である、請求項1に記載の方法。
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US11/644,318 US7674622B2 (en) | 2006-12-22 | 2006-12-22 | Method for determination of nucleated red blood cells and leukocytes in a whole blood sample in an automated hematology analyzer |
US11/644,318 | 2006-12-22 | ||
PCT/US2007/086217 WO2008079599A1 (en) | 2006-12-22 | 2007-12-03 | Method for determination of nucleated red blood cells and leukocytes in a whole blood sample in an automated hematology analyzer |
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CN101750476B (zh) * | 2008-12-08 | 2015-06-03 | 深圳迈瑞生物医疗电子股份有限公司 | 血液分析试剂及其使用方法 |
US8906309B2 (en) * | 2009-04-27 | 2014-12-09 | Abbott Laboratories | Method for discriminating red blood cells from white blood cells by using forward scattering from a laser in an automated hematology analyzer |
US8911669B2 (en) | 2009-08-24 | 2014-12-16 | Abbott Laboratories | Method for flagging a sample |
US8906308B2 (en) | 2010-01-15 | 2014-12-09 | Abbott Laboratories | Method for determining volume and hemoglobin content of individual red blood cells |
EP3246709A3 (en) | 2010-09-16 | 2018-02-14 | The General Hospital Corporation | Red blood cell dynamics for diagnosis |
US9005909B2 (en) | 2011-01-06 | 2015-04-14 | Rigel Pharmaceuticals, Inc. | Whole blood assay for measuring AMPK activation |
CN103492875B (zh) * | 2011-04-28 | 2016-03-09 | 希森美康株式会社 | 血液分析装置及血液分析方法 |
EP2705135B1 (en) | 2011-05-04 | 2019-10-30 | Abbott Laboratories | Basophil analysis method |
WO2012151103A2 (en) | 2011-05-04 | 2012-11-08 | Abbott Laboratories | Nucleated red blood cell analysis system and method |
WO2012151102A2 (en) | 2011-05-04 | 2012-11-08 | Abbott Laboratories | White blood cell analysis system and method |
CN104755905B (zh) * | 2012-12-31 | 2018-07-03 | 贝克曼考尔特公司 | 具有凝集块校准的血小板计数的系统和方法 |
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