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Licensed Unlicensed Requires Authentication Published by De Gruyter April 12, 2022

Importance of cerebrospinal fluid storage conditions for the Alzheimer’s disease diagnostics on an automated platform

Rosa Ferrer, Nuole Zhu, Javier Arranz, Inmaculada Porcel, Shaimaa El Bounasri, Oriol Sánchez, Soraya Torres, Josep Julve, Alberto Lleó, Francisco Blanco-Vaca, Daniel Alcolea and Mireia Tondo ORCID logo

Abstract

Objectives

Alzheimer’s disease (AD) is considered the most common cause of dementia in older people. Cerebrospinal fluid (CSF) Aβ1-42, Aβ1-40, total Tau (t-Tau), and phospho Tau (p-Tau) are important biomarkers for the diagnosis, however, they are highly dependent on the pre-analytical conditions. Our aim was to investigate the potential influence of different storage conditions on the simultaneous quantification of these biomarkers in a fully-automated platform to accommodate easier pre-analytical conditions for laboratories.

Methods

CSF samples were obtained from 11 consecutive patients. Aβ1-42, Aβ1-40, p-Tau, and t-Tau were quantified using the LUMIPULSE G600II automated platform.

Results

Temperature and storage days significantly influenced Aβ1-42 and Aβ1-40 with concentrations decreasing with days spent at 4 °C. The use of the Aβ1-42/Aβ1-40 ratio could partly compensate it. P-Tau and t-Tau were not affected by any of the tested storage conditions. For conditions involving storage at 4 °C, a correction factor of 1.081 can be applied. Diagnostic agreement was almost perfect in all conditions.

Conclusions

Cutoffs calculated in samples stored at −80 °C can be safely used in samples stored at −20 °C for 15–16 days or up to two days at RT and subsequent freezing at −80 °C. For samples stored at 4 °C, cutoffs would require applying a correction factor, allowing to work with the certainty of reaching the same clinical diagnosis.


Corresponding authors: Daniel Alcolea, Sant Pau Memory Unit, Department of Neurology, Hospital de la Santa Creu i Sant Pau, Biomedical Research Institute (IIB) Sant Pau, C/Sant Quintí 89, 08041, Barcelona, Spain; and Center of Biomedical Investigation Network for Neurodegenerative Diseases (CIBERNED), Madrid, Spain; and Mireia Tondo, Department of Biochemistry, Hospital de la Santa Creu i Sant Pau, Biomedical Research Institute (IIB) Sant Pau, Barcelona, Spain; Center of Biomedical Investigation Network for Diabetes and Metabolic Diseases (CIBERDEM), Madrid, Spain; and Comisión de Neuroquímica y Enfermedades Neurológicas, Sociedad Española de Medicina de Laboratorio, Barcelona, Spain, Phone: +34 93 5537358, Fax: +34 93 553787, E-mail:

Funding source: CIBERDEM

Funding source: CIBERNED

Funding source: Instituto de Salud Carlos III

Award Identifier / Grant number: PI21/00140

Award Identifier / Grant number: PI18/00435

Award Identifier / Grant number: INT19/00016

Award Identifier / Grant number: PI17/01896

Award Identifier / Grant number: AC19/00103

Funding source: Fondo Europeo de Desarrollo Regional

Funding source: Unión Europea

Funding source: Generalitat de Catalunya

Award Identifier / Grant number: 2017-SGR-547

Award Identifier / Grant number: SLT006/17/125

Award Identifier / Grant number: SLT002/16/408

Funding source: Marató TV3

Award Identifier / Grant number: 20142610

Acknowledgments

We thank all the participants of this study and all the members of the clinical and biochemical teams involved in the study.

  1. Research funding: This work was supported by CIBERDEM and CIBERNED and Instituto de Salud Carlos III (PI21/00140 to FB-V and MT, PI18/00435 and INT19/00016 to DA, PI17/01896 and AC19/00103 to AL), funded by Fondo Europeo de Desarrollo Regional (FEDER), Unión Europea, “Una manera de hacer Europa”. This work was also supported by Generalitat de Catalunya (2017-SGR-547, SLT006/17/125 to DA, SLT002/16/408 to AL) and “Marató TV3” foundation grants 20142610 to AL. We thank Fujirebio Europe NV for kindly providing the necessary reagents to perform the study.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent was obtained from all individuals included in this study.

  5. Ethical approval: All participants gave written informed consent before enrollment in accordance with the guidelines of the local Ethics Committee.

References

1. Cruz de Souza, L, Sarazin, M, Goetz, C, Dubois, B. Clinical investigations in primary care. Front Neurol Neurosci 2009;24:1–11.10.1159/000197897Search in Google Scholar

2. Jack, CR, Bennett, DA, Blennow, K, Carrillo, MC, Dunn, B, Haeberlein, SB, et al.. NIA-AA Research Framework: toward a biological definition of Alzheimer’s disease. Alzheimer’s Dementia 2018;14:535–62. https://doi.org/10.1016/j.jalz.2018.02.018.Search in Google Scholar

3. Dubois, B, Feldman, HH, Jacova, C, Hampel, H, Molinuevo, JL, Blennow, K, et al.. Advancing research diagnostic criteria for Alzheimer’s disease: the IWG-2 criteria. Lancet Neurol 2014;13:614–29. https://doi.org/10.1016/S1474-4422(14)70090-0.Search in Google Scholar

4. Lleó, A, Cavedo, E, Parnetti, L, Vanderstichele, H, Herukka, SK, Andreasen, N, et al.. Cerebrospinal fluid biomarkers in trials for Alzheimer and Parkinson diseases. Nat Rev Neurol 2015;11:41–55. https://doi.org/10.1038/nrneurol.2014.232.Search in Google Scholar PubMed

5. van der Kant, R, Goldstein, LSB, Ossenkoppele, R. Amyloid-β-independent regulators of tau pathology in Alzheimer disease. Nat Rev Neurosci 2020;21:21–35. https://doi.org/10.1038/s41583-019-0240-3.Search in Google Scholar PubMed

6. Blennow, K. Dementia in 2010: paving the way for Alzheimer disease drug development. Nat Rev Neurol 2011;7:65–6. https://doi.org/10.1038/nrneurol.2010.214.Search in Google Scholar PubMed

7. Lewczuk, P, Riederer, P, O’Bryant, SE, Verbeek, MM, Dubois, B, Visser, PJ, et al.. Cerebrospinal fluid and blood biomarkers for neurodegenerative dementias: an update of the Consensus of the Task Force on Biological Markers in Psychiatry of the World Federation of Societies of Biological Psychiatry. World J Biol Psychiatr : Off J World Fed Soc Biol Psychiatr 2018;19:244. https://doi.org/10.1080/15622975.2017.1375556.Search in Google Scholar PubMed PubMed Central

8. Hansson, O, Seibyl, J, Stomrud, E, Zetterberg, H, Trojanowski, JQ, Bittner, T, et al.. CSF biomarkers of Alzheimer’s disease concord with amyloid-β PET and predict clinical progression: a study of fully automated immunoassays in BioFINDER and ADNI cohorts. Alzheimer’s Dementia 2018;14:1470–81. https://doi.org/10.1016/J.JALZ.2018.01.010.Search in Google Scholar PubMed PubMed Central

9. Lehmann, S, Yang, Z, Park, SA, Leitão, MJ, Baldeiras, I, Herukka, S-K, et al.. Chasing the effects of pre-analytical confounders – a multicenter study on CSF-AD biomarkers. Front Neurol 2015;8:153. https://doi.org/10.3389/fneur.2015.00153.Search in Google Scholar PubMed PubMed Central

10. Bjerke, M, Portelius, E, Minthon, L, Wallin, A, Anckarster, H, Anckarster, R, et al.. Confounding factors influencing amyloid beta concentration in cerebrospinal fluid. Int J Alzheimer’s Dis 2010. https://doi.org/10.4061/2010/986310.Search in Google Scholar PubMed PubMed Central

11. Delaby, C, Muñoz, L, Torres, S, Nadal, A, le Bastard, N, Lehmann, S, et al.. Impact of CSF storage volume on the analysis of Alzheimer’s disease biomarkers on an automated platform. Clin Chim Acta 2019;490:98–101. https://doi.org/10.1016/J.CCA.2018.12.021.Search in Google Scholar

12. Kaiser, E, Schönknecht, P, Thomann, PA, Hunt, A, Schröder, J. Influence of delayed CSF storage on concentrations of phospho-tau protein (181), total tau protein and beta-amyloid (1–42). Neurosci Lett 2007;417:193–5. https://doi.org/10.1016/J.NEULET.2007.02.045.Search in Google Scholar PubMed

13. Lachno, DR, Romeo, MJ, Siemers, ER, Vanderstichele, H, Coart, E, Konrad, RJ, et al.. Supplementary data validation of ELISA methods for quantification of total tau and phosphorylated-tau 181 in human cerebrospinal fluid with measurement in specimens from two Alzheimer’s disease studies. Alzheimers Disease 2011;26:531–41.10.3233/JAD-2011-110296Search in Google Scholar PubMed

14. Zimmermann, R, Lelental, N, Ganslandt, O, Maler, JM, Kornhuber, J, Lewczuk, P. Preanalytical sample handling and sample stability testing for the neurochemical dementia diagnostics. J Alzheim Dis 2011;25:739–45. https://doi.org/10.3233/JAD-2011-110212.Search in Google Scholar PubMed

15. Janelidze, S, Stomrud, E, Brix, B, Hansson, O. Towards a unified protocol for handling of CSF before β-amyloid measurements. Alzheimer’s Res Ther 2019;11. https://doi.org/10.1186/s13195-019-0517-9.Search in Google Scholar PubMed PubMed Central

16. Hansson, O, Rutz, S, Zetterberg, H, Bauer, E, Hähl, T, Manuilova, E, et al.. Pre-analytical protocol for measuring Alzheimer’s disease biomarkers in fresh CSF. Alzheimer’s Dementia: Diagn Assess Dis Monit 2020;12. https://doi.org/10.1002/DAD2.12137.Search in Google Scholar PubMed PubMed Central

17. Schoonenboom, NSM, Mulder, C, Vanderstichele, H, van Elk, E-J, Kok, A, van Kamp, GJ, et al.. Effects of processing and storage conditions on amyloid (1-42) and tau concentrations in cerebrospinal fluid: implications for use in clinical practice. Clin Chem 2005;51:189–95. https://doi.org/10.1373/clinchem.2004.039735.Search in Google Scholar PubMed

18. Vanderstichele, H, de Vreese, K, Blennow, K, Andreasen, N, Sindic, C, Ivanoiu, A, et al.. Analytical performance and clinical utility of the INNOTEST® PHOSPHO-TAU(181P) assay for discrimination between Alzheimer’s disease and dementia with Lewy bodies. Clin Chem Lab Med 2006;44:1472–80. https://doi.org/10.1515/CCLM.2006.258.Search in Google Scholar PubMed

19. Sjögren, M, Davidsson, P, Tullberg, M, Minthon, L, Wallin, A, Wikkelso, C, et al.. Both total and phosphorylated tau are increased in Alzheimer’s disease. J Neurol Neurosurg Psychiatry 2001;70:624–30. https://doi.org/10.1136/jnnp.70.5.624.Search in Google Scholar PubMed PubMed Central

20. Blennow, K, Biscetti, L, Eusebi, P, Parnetti, L. Cerebrospinal fluid biomarkers in Alzheimer’s and Parkinson’s diseases-from pathophysiology to clinical practice. Mov Disord 2016;31:836–47. https://doi.org/10.1002/mds.26656.Search in Google Scholar PubMed

21. Bellomo, G, Cataldi, S, Paciotti, S, Paoletti, FP, Chiasserini, D, Parnetti, L. Measurement of CSF core Alzheimer disease biomarkers for routine clinical diagnosis: do fresh vs frozen samples differ? Alzheimers Res Ther 2020;12:121. https://doi.org/10.1186/s13195-020-00689-0.Search in Google Scholar PubMed PubMed Central

22. Bouwman, FH, van der Flier, WM, Schoonenboom, NSM, van Elk, EJ, Kok, A, Rijmen, F, et al.. Longitudinal changes of CSF biomarkers in memory clinic patients. Neurology 2007;69:1006–11.10.1212/01.wnl.0000271375.37131.04Search in Google Scholar PubMed

23. Alcolea, D, Clarimón, J, Carmona-Iragui, M, Illán-Gala, I, Morenas-Rodríguez, E, Barroeta, I, et al.. The Sant Pau Initiative on Neurodegeneration (SPIN) cohort: a data set for biomarker discovery and validation in neurodegenerative disorders. Alzheimer’s Dementia: Transl Res Clin Interv 2019;5:597–609. https://doi.org/10.1016/j.trci.2019.09.005.Search in Google Scholar PubMed PubMed Central

24. Alcolea, D, Pegueroles, J, Mu∼ Noz, L, Camacho, V, Opez-Mora, DL, Fern Andez-Le On, A, et al.. Agreement of amyloid PET and CSF biomarkers for Alzheimer’s disease on Lumipulse. Ann Clin Transl Neurol 2019;6:1815–24. https://doi.org/10.1002/acn3.50873.Search in Google Scholar PubMed PubMed Central

25. Schipke, CG, Jessen, F, Teipel, S, Luckhaus, C, Wiltfang, J, Esselmann, H, et al.. Long-term stability of Alzheimer’s disease biomarker proteins in cerebrospinal fluid. J Alzheim Dis 2011;26:255–62. https://doi.org/10.3233/JAD-2011-110329.Search in Google Scholar PubMed

Received: 2022-02-15
Accepted: 2022-03-24
Published Online: 2022-04-12
Published in Print: 2022-06-27

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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