A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking

Abstract. Progranulin (PGRN) levels in blood and cerebrospinal fluid (CSF) are increasingly studied as potential markers for neurodegenerative disorders. We aimed to 1) characterize two commercially available PGRN ELISAs on several assay validation parameters, 2) assess the stability of PGRN in serum and CSF under pre-analytical conditions, and 3) compare stability in the two assays. Intra-and inter-assay variation, inter-lot variation, linearity, lower limit of detection, and kit correlations were assessed for the Adipogen and R&D PGRN ELISA kits. Blood and serum samples were experimentally exposed to ≤9 freeze/thaw cycles, delayed processing for ≤24 h at room temperature and 4• C, and to temperature stability tests for ≤3 weeks at -20• C, 4• C, room temperature, and 37 • C. Both commercial PGRN ELISA kits showed acceptable ranges for intra-and inter-assay variation, where the R&D kit performed more accurate than the Adipogen kit, especially for inter-assay variation (intra-assay serum: 6.7 and 8.3%, respectively; inter-assay serum: 9.2 and 21.0%; intra-assay CSF: 3.6 and 12.0%; inter-assay CSF: 16.0 and 44.5%). Absolute serum PGRN concentrations were 1.9-fold higher in Adipogen than R&D (p < 0.001) and strongly correlated between both kits (ρ = 0.86, p < 0.0001) and CSF PGRN levels were on the borderline of detection in both kits. PGRN was typically stable under all pre-analytical conditions addressed, although two weeks at 37• C resulted in decreased PGRN concentrations in CSF, only when using the Adipogen kit. These results support further examination of PGRN as a potential marker in neurodegenerative diseases, since PGRN is stable in serum and CSF and can be measured using ELISA kits from several providers.

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“…2-6 ml clotted blood was processed and stored according to international consensus standard operation procedures [27].…”

Section: General Processing and Analysis Procedures Of Human Blood Anmentioning

confidence: 99%

Stability of Progranulin Under Pre-Analytical Conditions in Serum and Cerebrospinal Fluid

Willemse

Durieux-Lu

Flier

et al. 2016

JAD

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“…Lumbar CSF and paired blood samples were collected, centrifuged, aliquoted in 1 mL polypropylene tubes and stored on average within 1.5 h of sampling at −80 °C in accordance with established guidelines until analysis 34 . The specimens were thawed just prior to the measurements.…”

Section: Participantsmentioning

confidence: 99%

Neurofilaments and tau proteins in cerebrospinal fluid and serum in dementias and neuroinflammation

Fialová¹,

Bartoš²,

Švarcová³

2017

BIOMED PAP

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Aims. The aim of this study was to evaluate the diagnostic potential of cerebrospinal fluid (CSF) and serum levels of neurocytoskeletal proteins and their ratios for the diagnosis of dementias and to assess the differences in neurocytoskeletal proteins between neurodegeneration and neuroinflammation. Methods. CSF and serum levels of neurofilament light subunits (NFL) and neurofilament heavy subunits (NFH) as well as CSF levels of total tau (t-tau) and phosphorylated tau (p-tau) proteins were determined using ELISA in 20 Alzheimer's disease patients (AD group), 13 patients with other dementias (OD group), 17 patients with inflammatory aseptic neuro-infections (IP), and 20 aged-matched cognitively normal elderly persons (NC group). Results. The ratio CSF p-tau/t-tau was significantly higher in the NC group than that in the AD or OD groups (P<0.0005 for each group). The CSF NFH/p-tau and CSF NFL/p-tau ratios were significantly lower in AD patients than OD patients (P≤0.003). Serum and CSF NFL and CSF NFH levels were significantly higher in OD patients than AD patients (P≤0.03).The lowest values of the CSF NFL/NFH ratio were found in the IP group and they significantly differed from those in normal controls (P<0.0001) and any dementia group (IP vs. AD P<0.0001; IP vs. OD P=0.03). Conclusion. CSF tau proteins and their index differentiated between AD or OD patients and cognitively normal subjects, while CSF levels of neurofilaments expressed as their index seem to contribute to the discrimination between patients with neuroinflammation and normal controls or AD patients.

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“…It is however recommended to freeze and store Aβ standards and CSF at -80 °C to avoid stability issues [35][36][37]. Addition of protease inhibitors could improve protein stability upon storage but add an additional complexity to samples that could affect their commutability [37].…”

Section: Csf Sample Storagementioning

confidence: 99%

Quantitative detection of amyloid-β peptides by mass spectrometry: state of the art and clinical applications

Bros¹,

Delatour

Vialaret

et al. 2015

Clinical Chemistry and Laboratory Medicine (CCLM)

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Alzheimer's disease (AD) is the most common form of dementia in humans, and a major public health concern with 35 million of patients worldwide. Cerebrospinal fluid (CSF) biomarkers being early diagnostic indicators of AD, it is essential to use the most efficient analytical methods to detect and quantify them accurately. These biomarkers, and more specifically amyloid-β (Aβ) peptides, are measured in routine clinical practice using immunoassays. However, there are several limits to this immunodetection in terms of specificity and multiplexing of the multiple isoforms of the Aβ peptides. To overcome these issues, the quantification of these analytes by mass spectrometry (MS) represents an interesting alternative, and several assays have been described over the past years. This article reviews the different Aβ peptides quantitative MS-based approaches published so far, compares their pre-analytical phase, and the different quantitative strategies implemented that might be suitable for clinical applications.

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A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking

Cited by 642 publications

References 48 publications

Stability of Progranulin Under Pre-Analytical Conditions in Serum and Cerebrospinal Fluid

Stability of Progranulin Under Pre-Analytical Conditions in Serum and Cerebrospinal Fluid

Neurofilaments and tau proteins in cerebrospinal fluid and serum in dementias and neuroinflammation

Quantitative detection of amyloid-β peptides by mass spectrometry: state of the art and clinical applications

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