Cell analysis in cerebrospinal fluid (CSF) using Sysmex® hematology analyzers XT-4000i and XE-5000: Evaluation with CSF controls of the Joint German Society for Clinical Chemistry and Laboratory Medicine (DGKL)

被引:18
作者
Kleine, Tilmann O. [1 ]
Nebe, Carl Thomas [2 ]
Loewer, Christa [1 ]
Geilenkeuser, Wolf-Jochen [3 ]
Dorn-Beineke, Alexandra [4 ]
机构
[1] Univ Klinikum Giessen & Marburg, Inst Lab Med & Pathobiochem, Referenzlab Liquordiagnost, D-35043 Marburg, Germany
[2] Inst Immunol & Genet, ONKOLOGIKUM Kaiserslautern Speziallab Hamatol & O, D-67655 Kaiserslautern, Germany
[3] Referenzinst Bioanalyt, D-53175 Bonn, Germany
[4] Dr Horst Schmidt Kliniken GmbH, HSK, Inst Lab Diagnost & Hyg, D-65199 Wiesbaden, Germany
关键词
cerebrospinal fluid cell analysis; chamber cell counting; flow cytometry; hematology analyzers Sysmex (R) XT-4000i; XE-5000; LINEAR-REGRESSION PROCEDURES; LEUKOCYTES; EXPRESSION; ANTIGENS; MODE;
D O I
10.1002/cyto.a.22014
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
In cerebrospinal fluid (CSF) analysis, hematology analyzers (HAs) Sysmex (R) XT-4000i and XE-5000, equipped with flow cytometry (FCM), were used to count cells and differentiate leukocytes into mononuclear and polymorphonuclear cells (MNCs, PMCs) applying body fluid mode. FCM was evaluated with 20 DGKL CSF controls containing viable human leukocytes and erythrocytes. HA values were compared with reference values by Passing/Bablok regression analysis to reveal conformity. Conformity of white blood cells (WBCs) was obtained with native leukocytes, counted in calibrated FuchsRosenthal chamber as reference; red blood cell counts proved inaccurate. CV <40% with WBC counts <20 per mu L impairs accuracy. Reference WBC differentiation was assayed using FACS Canto II (TM) and FC-500 SN with anti-CD45, anti-CD14, anti-CD16, anti-CD16/56 [Becton Dickinson (BD); Beckman Coulter (BC)]. BD FACS lysing solution (R)-no-wash-procedure was applied. BC pretreatment with Versalyse lysing solution was not recommended. MNCs (lymphocytes + monocytes) were significantly lower (similar to 14%) on both HAs; PMCs (granulocytes or sum of neutrophils + eosinophils + basophils: range 186 M/L) were significantly higher (similar to 2.2-fold). WBC HA differentiation is not reliable because MNC/PMC differentiation yielded lower and higher values than FACSFCM references, respectively. This is attributed to incorrect discrimination of leukocytes with rounded/nonrounded nuclei; adding leukocytes with nonrounded nuclei to too low HA MNCs (about 40% not-activated) yielded P/B conformity; subtraction of leukocytes with nonrounded nuclei from elevated HA PMCs showed conformity (about 85% activated). Nucleus/activation state of leukocytes was assessed using microhistology. Sysmex XT-4000i and XE-5000 HAs systems are inappropriate for complete CSF cell analysis. (C) 2012 International Society for Advancement of Cytometry
引用
收藏
页码:255 / 264
页数:10
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