- 作者：Marleen J.A. Koel-Simmelink ; Anke Vennegoor ; Joep Killestein ; Marinus A. Blankenstein ; Niklas Norgren ; Carsten Korth ; Charlotte E. Teunissen
- 关键词：Nf ; neurofilament ; NfL ; neurofilament light ; NfH ; neurofilament heavy ; ELISA ; Enzyme Linked Immuno Sorbent Assay ; CSF ; cerebrospinal fluid ; MS ; multiple sclerosis ; CIS ; clinically isolated syndrome ; LQC ; low quality control ; HQC ; high quality control ; LOL ; Limit of linearity ; LOD ; limit of detection ; SD ; standard deviation ; ALS ; amyotrophic lateral sclerosis ; AD ; Alzheimer disease ; FTD ; frontotemporal dementia ; LBD ; Lewy bodies disease ; LP ; lumbar puncture
- 刊名：Journal of Immunological Methods
- 出版年：15 January, 2014
- 年：2014
- 卷：402
- 期：1-2
- 页码：43-49
- 全文大小：376 K
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 NfL 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 鈭?#xA0;20 掳C, 4 掳C, and room temperature. A decrease in concentration was observed during storage of both neurofilament proteins up to 21 days at 37 掳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.