Throughout the past ten years, metal-organic frameworks (MOFs) represent a class of coordination polymers characterized by high porosity and structural flexibility, and are increasingly used in biochemical analysis. However, largely due to the extremely-low content of biomolecules, the transferability and support between MOFs material and sensing substrate are crucial for sensing sensitivity and stability, which directly affect their practical applications. Rapid in-situ MOF synthesis strategy is obviously very important, which usually involves the direct growth of MOFs onto the sensing substrate in a "one-pot" process for simple, fast, portable and ideally large-scale preparation. This review focuses on the most current developments in the field of using MOFs produced quickly in-situ for highly sensitive detection of tiny amounts of protein, glucose, dopamine, ascorbic acid and uric acid by electrochemical, fluorescence, Raman, colorimetric sensing and microfluidics techniques.