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Effects of Processing and Storage Conditions on Amyloid ß (1–42) and Tau Concentrations in Cerebrospinal Fluid: Implications for Use in Clinical Practice

¹ Alzheimer Center and Department of Neurology, and ² Department of Clinical Chemistry, VU University Medical Center, Amsterdam, The Netherlands.
³ Innogenetics NV, Ghent, Belgium.

^aAddress correspondence to this author at: Departments of Neurology and Clinical Chemistry, VU University Medical Center, Postbus 7057, 1081 HV Amsterdam, The Netherlands. Fax 31-204440715; e-mail niki.schoonenboom{at}vumc.nl

Background: Reported concentrations of amyloid ß (1–42) (Aß42) and tau in cerebrospinal fluid (CSF) differ among reports. We investigated the effects of storage temperature, repeated freeze/thaw cycles, and centrifugation on the concentrations of Aß42 and tau in CSF.

Methods: Stability of samples stored at –80 °C was determined by use of an accelerated stability testing protocol according to the Arrhenius equation. Aß42 and tau concentrations were measured in CSF samples stored at 4, 18, 37, and –80 °C. Relative CSF concentrations (%) of the biomarkers after one freeze/thaw cycle were compared with those after two, three, four, five, and six freeze/thaw cycles. In addition, relative Aß42 and tau concentrations in samples not centrifuged were compared with samples centrifuged after 1, 4, 48, and 72 h.

Results: Aß42 and tau concentrations were stable in CSF when stored for a long period at –80 °C. CSF Aß42 decreased by 20% during the first 2 days at 4, 18, and 37 °C compared with –80 °C. CSF tau decreased after storage for 12 days at 37 °C. After three freeze/thaw cycles, CSF Aß42 decreased 20%. CSF tau was stable during six freeze/thaw cycles. Centrifugation did not influence the biomarker concentrations.

Conclusions: Repeated freeze/thaw cycles and storage at 4, 18, and 37 °C influence the quantitative result of the Aß42 test. Preferably, samples should be stored at –80 °C immediately after collection.

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