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[Display omitted] • The V O -NCO NWs incorporated low-valence redox species was achieved. • Abundant reactive redox species and enhanced intrinsic electrical conductivity in V O -NCOs NWs endow excellent catalytic activity. • A FMTE assembled with V O -NCOs NWs exhibits high-efficient glucose detection in sweat. NiCo 2 O 4 (NCO), a typical spinel-type transition metal oxide, emerges as a promising candidate for non-electroenzymatic glucose sensors because of its stable structure and low overpotential. However, the practical applications of NCO are hindered by its limited intrinsic activity for glucose detection. Herein, we intentionally incorporated low-valence redox species into NCO nanowires by modulating the oxygen vacancies (V O -NCO NWs), thereby facilitating glucose oxidation reactivity. The concentration of low-valence species increases with the generated oxygen vacancies, which enriches the reactive redox species and enhances the intrinsic electrical conductivity, thereby improving the catalytic activity. Consequently, the optimized V O -NCO NWs exhibit highly efficient glucose detection with high sensitivity up to 19.65 mA·mM−1·cm−2, which is 4.6 times higher than that of the pristine NCO NWs, as well as a low limit of detection (LOD) of 0.15 μM. Furthermore, a flexible miniaturized three electrode (FMTE) assembled with optimized V O -NCO NWs provides a high sensitivity of 15.89 mA·mM−1·cm−2 and a low LOD of 0.18 μM in synthetic sweat. Under various bending conditions, the FMTE demonstrates good mechanical flexibility, with no discernible alterations in the response current. These findings provide a promising protocol for the dynamic monitoring of sweat composition to reveal human physiological health status. [ABSTRACT FROM AUTHOR]