title: Use of TransFix™ cerebrospinal fluid storage tubes prevents cellular loss and enhances flow cytometric detection of malignant hematological cells after 18 hours of storage.
abstract: Flow cytometry is a sensitive method for detection of leptomeningeal localizations of hematological malignancies (LHM) in cerebrospinal fluid (CSF). Rapid processing of CSF is needed, as leukocyte numbers appear to decline quickly after lumbar puncture. The cell-stabilizing agent TransFix™ may enhance the detection of LHM in CSF by preventing cellular loss. To study the effects of TransFix on leukocyte numbers and the detection of LHM, we prospectively collected 99 CSF samples from patients with suspected or proven LHM in tubes with (i) TransFix; (ii) serum-containing medium; and (iii) no cell-stabilizing agents (native CSF). Presence of LHM and absolute leukocyte numbers were determined by flow cytometry after 30 minutes and 18 hours of storage. Leukocyte numbers in TransFix-stabilized CSF were higher than in the corresponding native samples at both time points (1.4× and 2.3× respectively, P < 0.0001 on each occasion). After 18 hours of storage, TransFix enhanced the detection of LHM in CSF. In all discordant paired observations (13/99, P = 0.005), the level of suspicion (classified as positive, suspicious, or negative) in CSF with TransFix was higher than in native CSF. We conclude that the use of TransFix-containing CSF storage tubes prevents cellular loss and enhances flow cytometric detection of LHM after 18 hours of storage.
authors.affiliation: Department of Medical Oncology, Erasmus University Medical Center/Daniel Den Hoed Cancer Center, Groene Hilledijk 301, 3075 EA Rotterdam, The Netherlands; Department of Neurology, Erasmus University Medical Center, Rotterdam's-Gravendijkwal 230, 3015 CE Rotterdam, The Netherlands.
authors.forename: Adriaan H
authors.initials: AH
authors.lastname: de Jongste
authors.forename: Jaco
authors.initials: J
authors.lastname: Kraan
authors.forename: Patricia D
authors.initials: PD
authors.lastname: van den Broek
authors.forename: Rik A
authors.initials: RA
authors.lastname: Brooimans
authors.forename: Jacoline E
authors.initials: JE
authors.lastname: Bromberg
authors.forename: Kees A
authors.initials: KA
authors.lastname: van Montfort
authors.forename: Peter A Sillevis
authors.initials: PA
authors.lastname: Smitt
authors.forename: Jan W
authors.initials: JW
authors.lastname: Gratama
mesh_terms: D000328:Adult; D000368:Aged; D002452:Cell Count; D002469:Cell Separation; D005260:Female; D005434:Flow Cytometry; D019337:Hematologic Neoplasms / Q000134:cerebrospinal fluid* / Q000175:diagnosis / Q000473:pathology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods*; D013997:Time Factors; D055815:Young Adult
keywords: FACSCanto cytofluorometric analysis; cerebrospinal fluid; hematological malignancies; leptomeningeal localization; lymphoma