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Highlights 1. We developed a biochip for rapid assay based on the magneto-optical Cotton–Mouton (CM) effect of γ-Fe2O3@Au core/shell magnetic nanoparticles. 2. The easy and quick assay for detection of the spike glycoprotein S of SARS-CoV-2 was demonstrated, and whole process takes approximately 50 min including 3 min for the CM rotation measurement with the detection limit of 0.27 ng/mL (3.4 pM). 3. This magneto-optical biochip we proposed can be easily modified to use as assays for other diseases.