We report a high-performance cholesterol biosensor using low-temperature solution grown ZnO hollow nanospheres (ZHNSs). The fabricated cholesterol biosensor showed a rapid response time of ∼2 s, a detection limit of 0.4 mM, a wide linear detection range (0.2–15.6 mM), and high sensitivity (99.8 μA mM⁻¹ cm⁻²). Owing to good selectivity and long-term stability, the biosensor manifests a cost-effective and highly potential platform for cholesterol detection.