Immunomagnetic reduction (IMR), an ultra-sensitive analytical assay, can reliably assay ultra-low concentrations of Aβ and tau biomarkers, including Aβ1–40, Aβ1–42, total tau (t-Tau), and p-Tau181 (Tsai et al., 2019). The levels of plasma Aβ1–40, Aβ1–42, t-Tau, and p-Tau181 were assessed using IMR kits (MF-AB0-0060, MF-AB2-0060, MF-TAU-0060, and MF-PT1-0060, MagQu Co., New Taipei City, Taiwan). For each assay, 40 μL (Aβ1–40, t-Tau, and p-Tau181) or 60 μL (Aβ1–42) of plasma was mixed with 80 or 60 μL of reagent, respectively. Each reported biomarker concentration represented the average of 2 duplicated measurements. An IMR analyzer (XacPro-S, MagQu Co., New Taipei City, Taiwan) was used for all assays. The measured biomarker concentrations ranged from 0.17 to 1,000 pg/mL for Aβ1–40, 0.77 to 30,000 pg/mL for Aβ1–42, 0.026 to 3,000 pg/mL for t-Tau, and 0.0196 to 1,000 pg/mL for p-Tau181. Intra-assay or inter-assay coefficients of variations using IMR ranged from 7 to 10% and from 10 to 15% for high-concentration and low-concentration quality control samples of Aβ1–40, Aβ1–42, t-Tau, or p-Tau181, respectively. For each biomarker, two batches of reagents were used, and the quality of each batch of reagents was well-controlled by monitoring the particle size, particle concentration, and bioactivity. The variation in these reagent properties between batches was lower than 10%.

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