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Recommendations for cerebrospinal fluid collection for the analysis by ELISA of neurogranin trunc P75, α-synuclein, and total tau in combination with Aβ(1-42)/Aβ(1-40).

Abstract：

BACKGROUND:The pathophysiology of neurodegeneration is complex. Its diagnosis requires an early identification of sequential changes in several hallmarks in the brains of affected subjects. The presence of brain pathology can be visualized in the cerebrospinal fluid (CSF) by protein profiling. It is clear that the field of Alzheimer's disease (AD) will benefit from an integration of algorithms including CSF concentrations of individual proteins, especially as an aid in clinical decision-making or to improve patient enrolment in clinical trials. The protein profiling approach requires standard operating procedures for collection and storage of CSF which must be easy to integrate into a routine clinical lab environment. Our study provides recommendations for analysis of neurogranin trunc P75, α-synuclein, and tau, in combination with the ratio of β-amyloid Aβ(1-42)/Aβ(1-40). METHODS:Protocols for CSF collection were compared with CSF derived from subjects with normal pressure hydrocephalus (n = 19). Variables included recipient type (collection, storage), tube volume, and addition of detergents at the time of collection. CSF biomarker analysis was performed with enzyme-linked immunosorbent assays (ELISAs). Data were analyzed with linear repeated measures and mixed effects models. RESULTS:Adsorption to recipients is lower for neurogranin trunc P75, α-synuclein, and tau (<10%), as compared to Aβ(1-42). For neurogranin trunc P75 and total tau, there is still an effect on analyte concentrations as a function of the tube volume. Protocol-related differences for Aβ(1-42) can be normalized at the (pre-)analytical level using the ratio Aβ(1-42)/Aβ(1-40), but not by using the ratio Aβ(1-42)/tau. The addition of detergent at the time of collection eliminates differences due to adsorption. CONCLUSIONS:Our study recommends the use of low protein binding tubes for quantification in CSF (without additives) of all relevant CSF biomarkers. Pre-analytical factors have less effect on α-synuclein, neurogranin trunc P75, and total tau, as compared to Aβ(1-42). The ratio of Aβ(1-42)/Aβ(1-40), but not Aβ(1-42)/tau, can be used to adjust for pre-analytical differences in analyte concentrations. Our study does not recommend the inclusion of detergents at the time of collection of CSF. The present results provide an experimental basis for new recommendations for parallel analysis of several proteins using one protocol for collection and storage of CSF.

journal_name

Alzheimers Res Ther

journal_title

Alzheimer's research & therapy

authors

Vanderstichele H,Demeyer L,Janelidze S,Coart E,Stoops E,Mauroo K,Herbst V,François C,Hansson O

doi

10.1186/s13195-017-0265-7

subject

Has Abstract

pub_date

2017-06-06 00:00:00

pages

issue

issn

1758-9193

pii

10.1186/s13195-017-0265-7

journal_volume

pub_type

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Recommendations for cerebrospinal fluid collection for the analysis by ELISA of neurogranin trunc P75, α-synuclein, and total tau in combination with Aβ(1-42)/Aβ(1-40).