Objectives: Paratuberculosis (PTBC) is a chronic infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP), with an important economic impact in the dairy sector. This study aimed at evaluating and comparing the performance of two molecular diagnostic tests (PCR and qPCR) for the detection of MAP shedding in feces and milk of seropositive dairy cows across time.