Automated Flow Cytometric Analysis of Blood Cells in Cerebrospinal Fluid Analytic Performance

St Olaf's University Hospital, Trondheim, Norway.
American Journal of Clinical Pathology (Impact Factor: 2.51). 06/2004; 121(5):690-700. DOI: 10.1309/EKFW-9E3L-LFXE-15X9
Source: PubMed


We compared the performance of an automated method for obtaining RBC and WBC counts and WBC differential counts in cerebrospinal fluid (CSF) samples with the reference manual method. Results from 325 samples from 10 worldwide clinical sites were used to demonstrate the accuracy, precision, and linearity of the method. Accuracy statistics for absolute cell counts showed a high correlation between methods, with correlation coefficients for all reportable absolute counts greater than 0.9. Linearity results demonstrated that the method provides accurate results throughout the reportable ranges, including clinical decision points for WBCs of 0 to 10/microL. Interassay precision and intra-assay precision for the ADVIA 120 (Bayer HealthCare, Tarrytown, NY) method were acceptable at all levels. The ADVIA 120 CSF Assay enumerates and differentiates cells via flow cytometry in a minimally diluted sample, improving the analysis of typically hypocellular CSF samples. Study results demonstrate that the automated ADVIA 120 CSF Assay is an acceptable alternative to the labor-intensive manual method.

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    • "One concern with this report was the extremely low detection rate of leptomeningeal disease by cytology, suggesting an unusual patient sample, suboptimal handling of CSF, or a high number of false positive results from flow cytometry. An additional benefit of flow cytometry is that automated methods are available that allow the rapid acquisition and processing of diagnostic data, meaning that its use may not lead to significant time expenditures from the clinicians or researchers if it is used to supplement conventional cytology routinely [30]. More recently, a larger study examining 123 patients with B-cell non-Hodgkin's lymphoma found that flow cytometric analysis using a 9-antibody panel detected 27 positive cases (22%) while cytology only detected 7 cases with 3 suspicious cases [31]. "
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    ABSTRACT: The spread of cancer into the central nervous system is a serious problem leading to neurological symptoms and rapid mortality. The current tools available for detecting the spread of cancer into the cerebrospinal fluid (CSF) are cytology, neurologic examination, and neuroimaging. All three of these methods can be applied in concert to reach a diagnosis, but they all suffer from a lack of sensitivity, leading to delays in treatment in many cases. An overview of research tools in the field of CSF cancer detection reveals a variety of promising technologies that can be used to answer questions about the biology of metastatic cancer and to develop more powerful clinical detection methods. Methods currently under investigation include new immunocytochemistry methods and flow cytometry for the in vitro detection of cells. Additionally, polymerase chain reaction, fluorescence in situ hybridization, capillary electrophoresis with laser-induced fluorescence, and mass spectrometry using matrix-assisted laser absorption-deionization time-of-flight and surface-enhanced laser desorption/ionization time-of-flight techniques are being tested for in vitro assessment of the non-cellular biomarkers in CSF. For in vivo detection of cancer in the CSF, research techniques include certain quantum dot platforms as well as magnetic iron oxide nanoparticles. As systemic therapies for cancer improve, the CNS is becoming a more common site of disease recurrence. This increases the importance of effective detection methods in the CSF, since early intervention can maximize therapeutic benefit. Furthermore, many cell-based detection methods can be combined with therapeutic agents to serve multiple medical functions through a common targeting system.
    Full-text · Article · Mar 2011 · Fluids and Barriers of the CNS
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    • "The automated counting is faster, easier as it does not require pretreatment of the samples and analytically more precise than manual counting and does not require a particularly qualified personnel. Several studies previously reported on automated counting of synovial fluid (Salinas et al., 1997; de Jonge et al., 2004), CSF (Ziebig, 2000; Aune et al., 2004), pleural fluid (Conner et al., 2003; De Jonge et al., 2006), ascitic/peritoneal fluid (Angeloni et al., 2003), and body fluids in general (Aulesa et al., 2003; Kresie et al., 2005). To compare the microscopic method with the automated XE-5000 method, we initiated a study and analyzed one hundred and seventy four (n = 174) body fluid samples including 81 ascitic fluids, 32 CSF, 26 pleural fluids, 18 synovial fluids, 13 peritoneal fluids, and 4 other types. "
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    ABSTRACT: We evaluated the performance of the automated body fluid mode of the Sysmex XE-5000 series automated haematology analyzer and compared the performance of the automated method for obtaining white blood cell (WBC), red blood cell (RBC) counts and WBC differential counts with microscopic method. One hundred and seventy-four samples were analysed: 81 ascitic fluid, 32 cerebrospinal fluid (CSF), 26 pleural fluid (PF), 18 synovial fluid (SF), 13 peritoneal fluid (PeF) and 4 other types. The agreement between the automated method and the manual reference showed high correlation, with Pearson correlation coefficients greater than 0.9 for all types of body fluids. We also demonstrate that the automated body fluid analysis on the XE-5000 is an acceptable alternative to the microscopic reference method as far as ascitic fluid, peritoneal dialysis fluid, SF or PF are concerned. Conversely, results for body fluid samples from oncology patients with leukaemia or tumours showed significant differences between both methods, as XE-5000 counted blast cells and neoplastic cells in mononuclear cell count. XE-5000 could represent an attractive method for the automated analysis of WBC, RBC, mononuclear cell count (MNC) and polymorphonuclear (PMN) cells of most body fluids. However, CSFs from patients with leukaemia or lymphoma should be processed with the microscopic reference method in order to detect abnormal leukaemic cells.
    Full-text · Article · Mar 2010 · International journal of laboratory hematology
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    • "Automated urine microscopy analyzers do not allow sensitive detection of bacteria for CSF applications [8] [9]. The application originally conceived for urinalysis flow cytometers for CSF applications heralded a new era [1] [3] [4]. Sysmex UF-100 (TOA Medical Electronics, Kobe, Japan), originally intended for urinalysis purposes, shows an apparent " noise " in the bacterial channel in microbiologically confirmed sterile CSF and continuous ambulatory peritoneal dialysis (CAPD) fluids [1]. "
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    ABSTRACT: Caution is of paramount importance in the interpretation of flow cytometric white blood cell counts in specimens with a high lymphocyte percentage as it is in the interpretation of bacterial counts in hemorrhagic and ventricular drainage CSF specimens. Recently, flow cytometry using a semiconductor laser along with forward and sideward scatter detection and also a dedicated bacterial channel has been developed (Sysmex UF-1000i). We explored the possible applications of this novel approach in the differential diagnosis of meningitis. Flow cytometry, microscopy and biochemical data of 161 CSF samples were analyzed. Microbiological analysis was performed in 53 suspected cases of meningitis. Good agreement for leukocytes was obtained between UF-1000i (rho=0.614) and UF-100 (rho=0.582) and microscopy and between both flow cytometers (rho=0.734). Lymphocytes were correctly classified as WBC by UF-1000i. Bacterial count on UF-1000i showed to be reliable in hemorrhagic samples and in samples collected by ventricular drainage for which interference by blood platelets and cell debris forms a known caveat on UF-100. Bacterial background signal in sterile CSF samples was absent on UF-1000i. One case of Cryptococcus neoformans meningitis, missed by UF-100 was properly detected by UF-1000i. Sysmex UF-1000i CSF flow cytometer offers the clinician an improved aid in directing the differential diagnosis of meningitis towards viral, bacterial or fungal causes.
    Full-text · Article · Jul 2008 · Clinica Chimica Acta
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