Format

Send to

Choose Destination
Alzheimers Res Ther. 2019 Jul 19;11(1):63. doi: 10.1186/s13195-019-0517-9.

Towards a unified protocol for handling of CSF before β-amyloid measurements.

Author information

1
Department of Clinical Sciences Malmö, Clinical Memory Research Unit, Lund University, Sölvegatan 19, BMC B11, 221 84, Lund, Sweden. shorena.janelidze@med.lu.se.
2
Department of Clinical Sciences Malmö, Clinical Memory Research Unit, Lund University, Sölvegatan 19, BMC B11, 221 84, Lund, Sweden.
3
Memory Clinic, Skåne University Hospital, Simrisbanvägen 14, SE-20502, Malmö, Sweden.
4
EUROIMMUN AG, 23560, Luebeck, Germany.
5
Department of Clinical Sciences Malmö, Clinical Memory Research Unit, Lund University, Sölvegatan 19, BMC B11, 221 84, Lund, Sweden. Oskar.Hansson@med.lu.se.
6
Memory Clinic, Skåne University Hospital, Simrisbanvägen 14, SE-20502, Malmö, Sweden. Oskar.Hansson@med.lu.se.

Abstract

BACKGROUND:

Widespread implementation of Alzheimer's disease biomarkers in routine clinical practice requires the establishment of standard operating procedures for pre-analytical handling of cerebrospinal fluid (CSF).

METHODS:

Here, CSF collection and storage protocols were optimized for measurements of β-amyloid (Aβ). We investigated the effects of (1) storage temperature, (2) storage time, (3) centrifugation, (4) sample mixing, (5) blood contamination, and (6) collection gradient on CSF levels of Aβ. For each study participant, we used fresh CSF directly collected into a protein low binding (LoB) tube that was analyzed within hours after lumbar puncture (LP) as standard of truth. Aβ42 and Aβ40 were measured in de-identified CSF samples using EUROIMMUN and Mesoscale discovery assays.

RESULTS:

CSF Aβ42 and Aβ40 were stable for at least 72 h at room temperature (RT), 1 week at 4 °C, and 2 weeks at - 20 °C and - 80 °C. Centrifugation of non-blood-contaminated CSF or mixing of samples before the analysis did not affect Aβ levels. Addition of 0.1-10% blood to CSF that was stored at RT without centrifugation led to a dose- and time-dependent decrease in Aβ42 and Aβ40, while Aβ42/Aβ40 did not change. The effects of blood contamination were mitigated by centrifugation and/or storage at 4 °C or - 20 °C. Aβ levels did not differ between the first to fourth 5-ml portions of CSF.

CONCLUSIONS:

CSF can be stored for up to 72 h at RT, 1 week at 4 °C, or at least 2 weeks at either - 20 °C or - 80 °C before Aβ measurements. Centrifugation of fresh non-blood-contaminated CSF after LP, or mixing before analysis, is not required. In case of visible blood contamination, centrifugation and storage at 4 °C or - 20 °C is recommended. After discarding the first 2 ml, any portion of up to 20 ml of CSF is suitable for Aβ analysis. These findings will be important for the development of a clinical routine protocol for pre-analytical handling of CSF.

KEYWORDS:

Alzheimer’s disease diagnosis; Biomarkers; Cerebrospinal fluid; Pre-analytical variables; β-Amyloid

Supplemental Content

Full text links

Icon for BioMed Central Icon for PubMed Central
Loading ...
Support Center