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Selective detection and quantification of biomarkers related to human diseases are essential for preventive healthcare. Surface-enhanced Raman scattering (SERS) spectroscopy is a powerful analytical tool offering high sensitivity. However, the success of this promising analytical tool relies on the ability to effectively fabricate SERS substrate. Herein we have demonstrated a plasmonic paper-based flexible substrate (PPFS) for SERS sensing. In situ growth of silver nanostructures (AgNS) on the paper-based substrate was achieved by using a simple one-step silver mirror reaction (SMR). FESEM and TEM results depicts that the increasing silver ion content influences the morphology (growth of multifacets), as well as size of AgNS. Further, the PPFS substrate was tested with Rhodamine-6G (Rh-6G) dye and an attomole sensitivity with a LOD of 4.54 x 10-18 M was achieved. Further, two biomarkers, lactic acid (LA) and uric acid (UA) were detected on the PPFS substrate, with μM and pM sensitivity, having LOD values of 0.6 x 10-6 and 0.3 x 10-12 M respectively. Above detection levels for UA on PPFS is two orders better than reported values, whereas for LA it is comparable with reported substrates. Finally, UA, LA and their mixtures were tested on PPFS and results compared with commercial substrate. The performance of PPFS were found better in all cases, thus, multifaceted AgNS paper based PPFS offers the potential to be used as a biosensor for detection of various biomarkers from body fluids, responsible for the detection of the critical disease for preventive health care.