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Pre-analytical stability of novel cerebrospinal fluid biomarkers

Journal article
Authors E. A. J. Willemse
Y. Vermeiren
M. S. Garcia-Ayllon
C. Bridel
P. P. De Deyn
S. Engelborghs
W. M. van der Flier
E. E. W. Jansen
I. B. Lopez-Font
V. Mendes
B. Manadas
N. de Roeck
J. Saez-Valero
E. A. Struys
E. Vanmechelen
Ulf Andreasson
C. E. Teunissen
Published in Clinica Chimica Acta
Volume 497
Pages 204-211
ISSN 0009-8981
Publication year 2019
Published at Institute of Neuroscience and Physiology
Pages 204-211
Language en
Keywords Assay validation, Biomarkers, Human cerebrospinal fluid, Neurodegenerative diseases, Pre-analytical stability
Subject categories Neurochemistry, Clinical chemistry


Stability of the cerebrospinal fluid (CSF) composition under different pre-analytical conditions is relevant for the diagnostic potential of biomarkers. Our aim was to examine the pre-analytical stability of promising CSF biomarkers that are currently evaluated for their discriminative use in various neurological diseases. Pooled CSF was aliquoted and experimentally exposed to delayed storage: 0, 1, 2, 4, 24, 72, or 168 h at 4 °C or room temperature (RT), or 1–4 months at −20 °C; or up to 7 freeze/thaw (f/t) cycles, before final storage at −80 °C. Eleven CSF biomarkers were screened using immunoassays, liquid chromatography, or enzymatic methods. Levels of neurogranin (truncP75), chitinase-3-like protein (YKL-40), beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), acetylcholinesterase (AChE) enzymatic activity, theobromine, secreted protein acidic and rich in cysteine-like 1 (SPARCL-1) and homovanillic acid (HVA) levels were not affected by the applied storage conditions. 3-Methoxy-4-hydroxyphenylglycol (MHPG) levels linearly and strongly decreased after 4 h at RT (−10%) or 24 h at 4 °C (−27%), and with 6% after every f/t cycle. 5-Methyltetrahydrofolate (5-MTHF) (−29% after 1 week at RT) and 5-hydroxyindoleacetic acid levels (5-HIAA) (−16% after 1 week at RT) were reduced and 3,4-dihydroxyphenylacetic acid (DOPAC) levels (+22% after 1 week at RT) increased, but only after >24 h at RT. Ten out of eleven potential CSF novel biomarkers showed very limited change under common storage and f/t conditions, suggesting that these CSF biomarkers can be trustfully tested under the pre-analytical conditions present across different cohorts. © 2019 Elsevier B.V.

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