CSF samples were obtained by LP at the L3/L4 or L4/L5 intervertebral level, centrifuged in case of blood contamination, divided into aliquots, and stored in polypropylene tubes at −80 °C until analysis. For the AD core biomarkers measurements, t-tau, p-tau, Aβ42, and Aβ40 were measured by automated chemiluminescent enzyme immunoassay on the Lumipulse G600II platform (Fujirebio, Gent, Belgium). The inter-assay coefficients of variation (CVs) were <8% for all biomarkers. Pathological values for defining the A/T status were determined using validated cutoff values [33 (link)]. More specifically, an Aβ42/Aβ40 ratio < 0.65 and a p-tau > 62 pg/mL supported the A+ and T+ statuses, respectively.
Commercially available ELISA kits were used to measure the NfL and 14-3-3 gamma isoform, as described [34 (link),35 (link)]. The GFAP concentrations were determined by running the commercially available GFAP Discovery Kit (Quanterix) on the SiMOA SR-X platform (Quanterix, Billerica, MA, USA). The intra-assay and the inter-assay CVs were respectively 7% and 15% for NfL, 6% and 13% for 14-3-3, and 8% for GFAP (only one plate was used). Eventually, all CSF samples from patients without autopsy examination, classified as probable sCJD or np-RPD, were tested by the second-generation prion RT-QuIC, as described [6 (link)].
Commercially available ELISA kits were used to measure the NfL and 14-3-3 gamma isoform, as described [34 (link),35 (link)]. The GFAP concentrations were determined by running the commercially available GFAP Discovery Kit (Quanterix) on the SiMOA SR-X platform (Quanterix, Billerica, MA, USA). The intra-assay and the inter-assay CVs were respectively 7% and 15% for NfL, 6% and 13% for 14-3-3, and 8% for GFAP (only one plate was used). Eventually, all CSF samples from patients without autopsy examination, classified as probable sCJD or np-RPD, were tested by the second-generation prion RT-QuIC, as described [6 (link)].
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