BDNF measurements in serum samples were performed using the BDNF Discovery Kit single-molecule array assay (Simoa®) from Quanterix (Cat no. 102039). Calibrators, participant samples, and two quality control samples per plate were measured in duplicate using a 2-step assay on a Simoa SR-X platform (Quanterix) according to manufactures protocols. Serum samples were diluted 1/1,000. The intra-assay concentration coefficient of variation was calculated for the duplicates of each participant sample and if they were more than 20% the sample was remeasured as per De Wolf et al. (2020 (link)). The intra-assay coefficient of variation for included sample measures was 7.2%. The median values for bi-participant replicates were then calculated and if the absolute difference between a replicate and the median value exceeded 10% of the median, the replicate was treated as an outlier and deleted from the statistical analysis. The mean of each biomarker concentration was then calculated for each participant sample.
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