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Licensed Unlicensed Requires Authentication Published by De Gruyter May 7, 2021

Cerebrospinal fluid hemoglobin levels as markers of blood contamination: relevance for α-synuclein measurement

  • Silvia Paciotti , Erik Stoops , Cindy François , Giovanni Bellomo , Paolo Eusebi , Hugo Vanderstichele , Davide Chiasserini ORCID logo EMAIL logo and Lucilla Parnetti ORCID logo EMAIL logo

Abstract

Objectives

Cerebrospinal fluid α-synuclein (CSF α-syn) represents a possible biomarker in Parkinson’s disease (PD) diagnosis. CSF blood contamination can introduce a bias in α-syn measurement. To date, CSF samples with a red blood cells (RBC) count >50 RBC × 106/L or haemoglobin (Hb) concentration >200 μg/L are excluded from biomarker studies. However, investigations for defining reliable cut-off values are missing.

Methods

We evaluated the effect of blood contamination on CSF α-syn measurement by a systematic approach in a cohort of 42 patients with different neurological conditions who underwent lumbar puncture (LP) for diagnostic reasons. CSF samples were spiked with whole blood and serially diluted to 800, 400, 200, 100, 75, 50, 25, 5, 0 RBC × 106/L. CSF α-syn and Hb levels were measured by ELISA.

Results

In neat CSF, the average concentration of α-syn was 1,936 ± 636 ng/L. This value increased gradually in spiked CSF samples, up to 4,817 ± 1,456 ng/L (+149% α-syn variation) in samples with 800 RBC × 106/L.

We established different cut-offs for discriminating samples with α-syn level above 5, 10, and 20% variation, corresponding to a Hb (RBC) concentration of 1,569 μg/L (37 RBC × 106/L), 2,082 μg/L (62 RBC × 106/L), and 3,118 μg/L (87 RBC × 106/L), respectively.

Conclusions

Our data show the high impact of CSF blood contamination on CSF α-syn levels, highlighting the measurement of Hb concentration as mandatory when assessing CSF α-syn. The thresholds we calculated are useful to classify CSF samples for blood contamination, considering as reliable only those showing a Hb concentration <1,569 μg/L.


Corresponding authors: Davide Chiasserini, Department of Medicine and Surgery, Section of Physiology and Biochemistry, University of Perugia, Sant’Andrea delle Fratte, 06132 Perugia, Italy, Phone: +39 0755858197, E-mail: ; and Prof. Lucilla Parnetti, Laboratory of Clinical Neurochemistry, Section of Neurology, Department of Medicine and Surgery, University of Perugia, Sant’Andrea delle Fratte, 06132 Perugia, Italy, Phone: +39 0755783545, E-mail:
Silvia Paciotti and Erik Stoops contributed equally to this work.

Award Identifier / Grant number: 10147

  1. Research funding: This work was supported by the Michael J. Fox Foundation (Grant ID: 10147).

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

  3. Competing interests: Erik Stoops and Cindy Francois are employees of ADx NeuroSciences. Hugo Vanderstichele is a co-founder of ADx NeuroSciences and a founder of Biomarkable bv. Lucilla Parnetti, Silvia Paciotti, Davide Chiasserini, Giovanni Bellomo and Paolo Eusebi have no conflict of interest to declare.

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

  5. Ethical approval: The study is in accordance with the Helsinki Declaration and it was approved by the local Ethics Committee (Comitato Etico Aziende Sanitarie Regione Umbria19369/AV).

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2020-1521).


Received: 2020-10-13
Accepted: 2021-04-26
Published Online: 2021-05-07
Published in Print: 2021-09-27

© 2021 Walter de Gruyter GmbH, Berlin/Boston

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