Procalcitonin (PCT) is a sensitive and specific biomarker for sepsis diagnosis. In this study, a novel ratio-typed electrochemical immunosensor was constructed for reliable and sensitive assay of PCT based on hierarchical PtCoIr nanowires/polyethylene polyamine-grafted-ferrocene (PtCoIr HNWs/PEPA-Fc) and porous SiO₂@Ag nanoparticles-toluidine blue (porous SiO₂@Ag NPs-TB). Importantly, the PtCoIr HNWs/PEPA-Fc was first modified on the sensing interface, which harvested stable and strong electrochemical signals for readout of Fc due to the enriched anchoring sites created by the PtCoIr HNWs. Meanwhile, porous SiO₂@Ag NPs-TB behaved as the label to conjugate with secondary antibody (Ab₂), which also provided another strong detection signals originated from TB confined in such porous structures. The resulting immunosensor displayed a measurable output of procalcitonin (PCT) in the dynamic scope of 0.001 ~ 100 ng mL⁻¹ with a low limit of detection (LOD) of 0.46 pg mL⁻¹ (S/N = 3). Moreover, we exploited this strategy for PCT assay in a diluted human serum sample with acceptable results, exhibiting promising applications in the clinical analysis.