Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 402, Issue 1-2, Pages 43-49Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2013.11.008
Keywords
Neurofilament heavy (NfH) protein; Neurofilament light (NfL) protein; Pre-analytical variables; Stability; NfH SinglePlex Luminex assay; Cerebrospinal fluid
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Funding
- EDAR
- Koningin Wilhelmina Fonds
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Background: Neurofilament (Nf) proteins have been shown to be promising biomarkers for monitoring and predicting disease progression for various neurological diseases. The aim of this study was to evaluate the effects of pre-analytical variables on the concentration of neurofilament heavy (NfH) and neurofilament light (NfL) proteins. Methods: For NfH an in-house newly-developed and validated SinglePlex Luminex assay was used; ELISA was used to analyze NfL. Results: For the NfL ELISA assay, the intra- and inter-assay variation was respectively, 1.5% and 16.7%. Analytical performance of the NfH SinglePlex Luminex assay in terms of sensitivity (6.6 pg/mL), recovery in cerebrospinal fluid (CSF) (between 90 and 104%), linearity (from 6.6-1250 pg/mL), and inter- and intra-assay variation (<8%) were good. Concentrations of both NI and NfH appeared not negatively affected by blood contamination, repeated freeze-thaw cycles (up to 4), delayed processing (up to 24 hours) and during long-term storage at -20 degrees C, 4 degrees C, and room temperature. A decrease in concentration was observed during storage of both neurofilament proteins up to 21 days at 37 degrees C, which was significant by day 5. Conclusions: The newly developed NfH SinglePlex Luminex assay has a good sensitivity and is robust. Moreover, both NfH and NfL are stable under the most prevalent pre-analytical variations. (C) 2013 Elsevier B.V. All rights reserved.
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