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Validation of a new assay for α-synuclein detection in cerebrospinal fluid

  • Marthe Gurine Førland , Annika Öhrfelt , Linn Silje Oftedal , Ole-Bjørn Tysnes , Jan Petter Larsen , Kaj Blennow , Henrik Zetterberg , Guido Alves and Johannes Lange ORCID logo EMAIL logo
Published/Copyright: July 30, 2016
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 55 Issue 2

Abstract

Background:

Abnormal α-synuclein aggregation and deposition is the pathological hallmark of Parkinson’s disease (PD) and dementia with Lewy bodies (DLB), but is also found in Alzheimer disease (AD). Therefore, there is a gaining interest in α-synuclein in cerebrospinal fluid (CSF) as potential biomarker for these neurodegenerative diseases. To broaden the available choices of α-synuclein measurement in CSF, we developed and validated a new assay for detecting total α-synuclein.

Methods:

This novel ELISA uses commercially available antibodies and is based on electrochemiluminescence technology. The assay protocol is straightforward, with short and simple incubation steps, and requires only small amounts of CSF. We validated this assay for precision, parallelism, dilution linearity, specificity, and spike recovery. We further compared it to the newly validated α-synuclein assay from BioLegend by analyzing a set of 50 CSF samples with both assays.

Results:

The new assay quantifies α-synuclein in CSF with a lower limit of detection of 36.3 pg/mL and shows no cross-reactivity with human β- and γ-synuclein. Results of dilution linearity, parallelism, spike recovery, and precision classify this assay as well suited for α-synuclein detection in human CSF samples.

Conclusions:

We present a novel assay based on freely available components to quantify total α-synuclein in CSF as an additional method for α-synuclein as a biomarker in neurodegenerative diseases. The assay convinces with its simple and convenient protocol paired with high sensitivity.

Keywords: α-synuclein; assay validation; biomarker; cerebrospinal fluid; ELISA
Corresponding author: Dr. Johannes Lange, The Norwegian Centre for Movement Disorders, Stavanger University Hospital, Postboks 8100, Stavanger 4068, Norway, Phone: +47-5151-5602, Fax: +47-5151-5515

Funding source: European Research Council

Award Identifier / Grant number: 681712

Funding statement: This study was supported by the Western Norway Regional Health Authority (grant nos. 911218, 911832 and 911698), the Norwegian Parkinson’s Disease Association, the Swedish Research Council (grant nos. 14002 and 2013-2546), the Torsten Söderberg Foundation, the Swedish Brain Foundation (grant no. FO2015-0021), the Knut and Alice Wallenberg Foundation, the European Research Council (grant no. 681712), and the Swedish Alzheimer Foundation (grant no. AF-553101).

Acknowledgments

The authors thank the CSF donors for participating in this study. We are grateful to Åsa Källén for her technical assistance. We would like to thank the Western Norway Regional Health Authority, and the Norwegian Parkinson’s Disease Association, the Swedish Research council, the Torsten Söderberg Foundation, the Swedish Brain Foundation, and the Swedish Alzheimer Foundation for support.

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

  2. Research funding: This study was supported by the Western Norway Regional Health Authority (grant nos. 911218, 911832 and 911698), the Norwegian Parkinson’s Disease Association, the Swedish Research Council (grant nos. 14002 and 2013-2546), the Torsten Söderberg Foundation, the Swedish Brain Foundation (grant no. FO2015-0021), the Knut and Alice Wallenberg Foundation, the European Research Council (grant no. 681712), and the Swedish Alzheimer Foundation (grant no. AF-553101).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organizations played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

  6. Financial disclosures/conflict of interest: None.

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Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2016-0409) offers supplementary material, available to authorized users.

Received: 2016-5-11
Accepted: 2016-7-4
Published Online: 2016-7-30
Published in Print: 2017-2-1

©2017 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2016-0409
Keywords for this article
α-synuclein; assay validation; biomarker; cerebrospinal fluid; ELISA

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Commutable samples with assigned target concentrations may help us harmonise general chemistry results
  4. Reviews
  5. Performance of point-of-care HbA1c test devices: implications for use in clinical practice – a systematic review and meta-analysis
  6. Cardiac troponins and mortality in type 1 and 2 myocardial infarction
  7. Opinion Paper
  8. Criteria for assigning laboratory measurands to models for analytical performance specifications defined in the 1st EFLM Strategic Conference
  9. Genetics and Molecular Diagnostics
  10. External quality assessment for human papillomavirus 16/18 DNA detection and genotyping in Shanghai, China
  11. General Clinical Chemistry and Laboratory Medicine
  12. Analytical performance of 17 general chemistry analytes across countries and across manufacturers in the INPUtS project of EQA organizers in Italy, the Netherlands, Portugal, United Kingdom and Spain
  13. Commutability of proficiency testing material containing tobramycin: a study within the framework of the Dutch Calibration 2.000 project
  14. Optimization and validation of moving average quality control procedures using bias detection curves and moving average validation charts
  15. Extending laboratory automation to the wards: effect of an innovative pneumatic tube system on diagnostic samples and transport time
  16. Smart management of sample dilution using an artificial neural network to achieve streamlined processes and saving resources: the automated nephelometric testing of serum free light chain as case study
  17. An integrated proteomic and peptidomic assessment of the normal human urinome
  18. An alternative inhibition method for determining cross-reactive allergens
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  20. Reference Values and Biological Variations
  21. Intra-individual variation of plasma trimethylamine-N-oxide (TMAO), betaine and choline over 1 year
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  25. Mid-regional pro-adrenomedullin (MR-proADM) and mid-regional pro-atrial natriuretic peptide (MR-proANP) in severe aortic valve stenosis: association with outcome after transcatheter aortic valve implantation (TAVI)
  26. Association between apolipoprotein E polymorphisms and premature coronary artery disease: a meta-analysis
  27. Urinary orosomucoid: a novel, early biomarker of sepsis with promising diagnostic performance
  28. Letters to the Editor
  29. CT or MRI
  30. Reply to: CT or MRI in the diagnosis of right lower quadrant abdominal pain?
  31. Quantification of daratumumab in the serum protein electrophoresis
  32. Response to: Interference of daratumumab on the serum protein electrophoresis
  33. Glycated albumin: correlation to HbA1c and preliminary reference interval evaluation
  34. Using “big data” to describe the effect of seasonal variation in thyroid-stimulating hormone
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