In order to satisfy requirement of point-of-care test(POCT) for rapid detection at primary-level clinics and personal users, a novel amperometric electrochemistry sensor detection system was developed and combined with pump-free microfluidic chip used for H2O2 and prostate cancer markers detection. Capillary force was utilized to modify polydimethylsiloxane channel for adding and mixing samples, a portable three-electrode detector was designed to control input signals and receive current signal on electrode sensor. This system was certificated by the portable detection circuit output voltage stability, and the error accuracy not exceed 1%. Meanwhile, detection of hydrogen peroxide showed good current dynamic response of concentration change. Prostate specific antigen was quantitatively detected with a limit of detection of 1ng/mL. To the best of our knowledge, this is the first time that PSA was detected using electrochemistry and pump-free microfluidic chip. It is a simple portable electrochemistry detection system based on pump-free microfluidic chip and expect to combine with different electrochemical sensors for rapid intelligent POCT detection.