Abstract: A surface plasmon resonance (SPR) sensor based on molecularly imprinted film (MIF) was proposed for the sensitive detection of progesterone. The MIF was synthesized by the method of thermo and photo polymerization on the sensor surface, which was subsequently removed to form the recognition cavities by the mixture of acetonitrile and acetic acid. The SPR sensor develops the angle shift by 1.15° (thermo polymerization) and 0.72° (photo polymerization) after washing progesterone from MIF and allowed detection of progesterone at concentration down to 10^-14 g/mL. The resonance angle changes show a good linearity versus minus logarithmic concentration of progesterone from 10^-14 g/mL to 10^-6 g/mL. A control experiment with the analogues estradiol, testosterone and testosterone propionate (10^-4 g/mL) shows a good selectivity of the MIF for the recognition of progesterone.