Rapid, accurate, and cost-effective strategies for the multiplexed and quantitative detection of multiple infectious disease biomarkers are still lack in hospital. Here, we introduce a multiplexed screen-printed electrode for the simultaneous detection of three infectious disease biomarkers: HBsAg, anti-HIV, and anti-TP. The electrode modified with in-situ deposition of gold nanoparticles (AuNPs) and MXene multilayered nanosheets. Comprehensive characterization confirms the morphology and electrochemical response of the AuNPs&MXene modified screen-printed electrode (AuNPs&MXene-SPE). AuNPs&MXene composite contributes to enhance sensitivity by maximizing surface area and improving electron transfer efficiency. The AuNPs&MXene-SPE exhibits outstanding detection performance for HBsAg, anti-HIV, and anti-TP. The AuNPs&MXene-SPE sensors achieve broad detection ranges (0.05–1000 ng/mL for HBsAg, 0.35–140 ng/mL for anti-HIV, and 0.25–100 ng/mL for anti-TP) accompanied by low limits of detection (0.01, 0.11, and 0.10 ng/mL, respectively). Applied to standard serum samples, the sensors showcase precision with relative standard deviations consistently below 5.0 %, affirming their suitability for complex solution environments. These sensors offer notable advantages, including cost-effectiveness, rapid response, high sensitivity, and specificity, eliminating the necessity for expensive equipment, which open avenues for the realization of point-of-care testing (POCT).
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