Venous blood was collected in a nonfasting state in lavender top EDTA tubes and placed on ice until processed. All blood was centrifuged less than 60 min from the time of blood draw, at 4°C at 3000 rpm for 10 min; plasma was isolated, and samples were stored in a −80°C freezer until batch assayed, blinded to all other study measures. Analyses were carried out using a proprietary single-molecule sensitive technology (31, 32). Briefly, plasma levels of tau were measured using the single-molecule enzyme-linked immunoarray (Simoa) method. A combination of monoclonal antibodies was used, where a subset of those antibodies detects tau by targeting the mid-region, while another subset is used for detection against the N terminus. This method provides molecular-level sensitivity while minimizing sample use and error levels.

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