Optical biosensors, especially those based on plasmonic structures, have emerged recently as a potential tool for disease diagnostics. Plasmonic biosensors have demonstrated impressive benefits for the label-free detection of trace biomarkers in human serum. However, widespread applications of these technologies are hindered because of their insufficient sensitivity, their relatively complex chemical immobilization processes, and the use of prism couplers. Accordingly, a sandwiched plasmon ruler (SW-PR) based on a Au nanohole array with ultrahigh sensitivity arising from the plasmonic coupling effect is developed. Highly confined surface charges caused by Bloch wave surface plasmon polarizations substantially increase the coupling efficiency. This platform exhibits thickness sensitivity as high as 61 nm nm and can detect at least 200 000-fold lower analyte concentrations than a nanowell sensing platform with the same wavelength shift. Additionally, the sandwiched plasmonic biosensor allows precise and label-free testing of clinical biomarkers, namely C-reactive protein and procalcitonin, in patient serum samples without requiring a sophisticated prism coupler, extra antibodies, or a chemical immobilization technique. This study yields new insight into the structural design of plasmon rulers and will open exciting avenues for disease diagnosis and therapy follow-up at the point-of-care.