Asunto(s)
Gránulos Citoplasmáticos/patología , Linfocitos/patología , Linfoma de Células T/patología , Médula Espinal/patología , Anciano , Encéfalo/patología , Caquexia , Trastornos del Conocimiento/etiología , Terapia Combinada , Encefalitis/patología , Resultado Fatal , Humanos , Inmunohistoquímica , Linfoma de Células T/líquido cefalorraquídeo , Linfoma de Células T/terapia , Imagen por Resonancia Magnética , Masculino , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunologíaRESUMEN
BACKGROUND: Manual cell counting in cerebrospinal fluid (CSF) is technique-dependent, time-consuming, and thus costly and prone to inter-operator variability and low precision. Flow cytometry (FCM) with fast hematology analyzers (HAs) appears to improve accuracy and precision of CSF cell analysis; rapid CSF cell analysis is especially needed in emergency laboratories. Ten external trials of the German Society for Clinical Chemistry and Laboratory Medicine evaluated FCM with Coulter (LH750, 755), Abbott CD3200, CD3500, CD3700, CD4000, Sapphire, ADVIA120 CSF assay, and Sysmex XE-2100 single platform analyzers. METHODS: CSF controls were produced using native blood leukocytes and erythrocytes, resembling CSF and thus rendering the trials feasible and allowing comparison with native manual counting in a Fuchs-Rosenthal chamber and FACScan-CD45-CD14 dual platform analysis, which was used as the reference method. Statistical evaluation was performed using Passing/Bablok regression analysis. RESULTS: Our evaluation revealed significant differences with respect to target values in leukocyte and erythrocyte counts, as well as leukocyte differentiation. These differences were attributed to inaccuracies produced by the HAs, due to blank correction in connection with impedance analysis, leukocyte loss, especially through monocyte injury due to the erythrocyte lysing agent, incomplete erythrocyte lysis, ADVIA cell sphering, cell differentiation using algorithms and peroxidase activity. Erythrocyte counting in the CSF controls was inaccurate with the Coulter and ADVIA analyzers. CONCLUSIONS: Evaluation of HAs by means of the CSF controls revealed inaccuracies in cell counting and leukocyte differentiation. Analyzer techniques, used for CSF cell assays, therefore need to be improved.
Asunto(s)
Líquido Cefalorraquídeo/citología , Citometría de Flujo/instrumentación , Recuento de Leucocitos/instrumentación , Leucocitos/citología , Diferenciación Celular , Técnicas de Laboratorio Clínico , Eritrocitos/citología , Alemania , Granulocitos/citología , Humanos , Linfocitos/citología , Monocitos/citología , Sociedades MédicasRESUMEN
Flow cytometry (FCM) is used with haematology analyzers (HAs) to count cells and differentiate leukocytes in cerebrospinal fluid (CSF). To evaluate the FCM techniques of HAs, 10 external DGKL trials with CSF controls were carried out in 2004 to 2008. Eight single platform HAs with and without CSF equipment were evaluated with living blood leukocytes and erythrocytes in CSF like DGKL controls: Coulter (LH750,755), Abbott CD3200, CD3500, CD3700, CD4000, Sapphire, ADVIA 120(R) CSF assay, and Sysmex XE-2100(R). Results were compared with visual counting of native cells in Fuchs-Rosenthal chamber, unstained, and absolute values of leukocyte differentiation, assayed by dual platform analysis with immune-FCM (FACSCalibur, CD45, CD14) and the chamber counts. Reference values X were compared with HA values Y by statistical evaluation with Passing/Bablock (P/B) linear regression analysis to reveal conformity of both methods. The HAs, studied, produced no valid results with DGKL CSF controls, because P/B regression revealed no conformity with the reference values due to:-blank problems with impedance analysis,-leukocyte loss with preanalytical erythrocyte lysis procedures, especially of monocytes,-inaccurate results with ADVIA cell sphering and cell differentiation with algorithms and enzyme activities (e.g., peroxidase). HA techniques have to be improved, e.g., using no erythrocyte lysis and CSF adequate techniques, to examine CSF samples precise and accurate.