Enzymes are considered as natural biocatalysts, which catalyze many biochemical reactions with good catalytic efficiency, biocompatibility, and substrate specificity. The intrinsic limitations of natural enzymes such as low stability, high cost, and storage difficulty have led to the introduction of artificial enzymes that imitate the activity of natural enzymes. With the rapid development of nanomaterials in the recent decade, novel enzyme-mimicking nanomaterials (nanozymes) have attracted considerable attention from researchers. Nanozymes are defined as a class of artificial nanomaterials possessing intrinsic enzymes-like activities, which have the advantages of simple preparation processes, low cost and some environmental tolerance. However, most of them are limited by their low activity and relatively poor stability, leading to many difficulties in the application of biochemical analysis. Among them, metal-organic framework nanozymes (MOFs) have demonstrated a wide range of uses because of their evident favorable circumstances, including the large surface area and porosity for functionalization, uniform active sites, high catalytic activity and stability, simple and controllable synthesis and low cost. In this review, we provide a summary of the clinical detection application of MOFs in nucleic acid, protein and small molecules based on their different activity classification (peroxidase, oxidase, catalase, superoxide dismutase, and hydrolase). Finally, we look forward to the opportunities and challenges that MOFs will face in clinical detection, promoting their clinical application transformation.