The facile preparation of highly sensitive electrochemical bioprobe based on lutetium phthalocyanine incorporated silica nanoparticles (SiO2(LuPc2)) grafted with Poly(vinyl alcohol-vinyl acetate) itaconic acid (PANI(PVIA)) doped polyaniline conducting nanobeads (SiO2(LuPc2)PANI(PVIA)-CNB) is reported. The preparation of CNB involves two stages (i) pristine synthesis of LuPc2 incorporated SiO2 and PANI(PVIA); (ii) covalent grafting of PANI(PVIA) onto the surface of SiO2(LuPc2). The morphology and other physico-chemical characteristics of CNB were investigated. The scanning electron microscopy images show that the average particle size of SiO2(LuPc2)PANI(PVIA)-CNB was between 180–220 nm. The amperometric measurements showed that the fabricated SiO2(LuPc2)PANI(PVIA)-CNB/GOx biosensor exhibited wide linear range (1–16 mM) detection of glucose with a low detection limit of 0.1 mM. SiO2(LuPc2)PANI(PVIA)-CNB/GOx biosensor exhibited high sensitivity (38.53 µA mM−1 cm−2) towards the detection of glucose under optimized conditions. Besides, the real (juice and serum) sample analysis based on a standard addition method and direct detection method showed high precision for measuring glucose at SiO2(LuPc2)PANI(PVIA)-CNB/GOx biosensor. The SiO2(LuPc2)PANI(PVIA)-CNB/GOx biosensor stored under refrigerated condition over a period of 45 days retains ~ 96.4% glucose response current.