Poly (acrylic acid) (PAA) nanocomposite hydrogel was fabricated as a sensing patch for non-invasive dual detection of urea in sweat. The hydrogel was prepared by γ-irradiation crosslinking of PAA solution incorporated with graphene oxide (GO) and cellulose nanofiber (CNF). With high water-sorption capacity and transparency, the hydrogel was suitable to accommodate coloring reagents and enzymes for colorimetric sensing of urea in sweat. The colorimetric sensor exhibited vivid color change towards the increase of urea concentration in a linear range of 40-80 mM covering a cut-off value (60 mM) for chronic kidney disease (CKD) indication. Furthermore, the hydrogel could be directly applied as a substrate for direct quantitation of urea in sweat by laser desorption ionization mass spectroscopy (LDI-MS). While CNF improved the mechanical properties of the hydrogel, GO played a key role in enhancing laser desorption ionization of urea in LDI-MS and increased the hydrogel functionalities. LDI-MS verified that GO/CNF/PAA hydrogel could act as a direct matrix for promoting urea ionization and these results corresponded well with the colorimetric sensor. Hence, this hydrogel patch might be a potential material to be applied in non-invasive and dual-detection of CKD in medical diagnosis.