300 mg of feces diluted in 1 mL of PBS, vortexed, centrifuged, supernatant added to a tube with zirconia beads and lysis-binding solution, vortexed, centrifuged, added to a plate with isopropanol, magnetic nucleic acid-binding beads and binding lysis enhancer, automated system used (MagMAX Express Magnetic Particle Processor)
Growth in Bactec 12B, 200 µL liquid culture medium added to 200 µL buffer RLT, transferred to a tube with 0.3 g zirconia/silica beads, disrupted in Fast Prep-24 bead beater for 60 s, and centrifuged, 100 µL added to a deep 96-well plate
MagMAX Total Nucleic Acid Isolation Kit; BioSprint 96 one-for-all vet kit (Qiagen) + MagMAX-96 automated magnetic processor; artificially contaminated only to set the Limit of Detection; auto = automatically processed; LOD = Limit of Detection; MAP = Mycobacterium avium subsp. paratuberculosis; n. a. = not available; PBS = phosphate-buffered saline; RLT = commercial name for the lysis buffer (Qiagen).