A microcolony‐immunoblot technique (MCIBI) was developed to directly enumerate, in less than 24 h, very low numbers ofListeria monocytogenes(8–12 colony forming units: CFU/g or mL) inoculated into foods. Four meat and poultry and two dairy products were artificially inoculated withL. monocytogenesV7 diluted and plated on Oxford agar medium. Each plate was overlaid with an Immobilon‐P membrane and incubated for 18–20 h at 37C. Blot‐transferred colonies on these membranes were probed with C11E9 monoclonal antibody (MAb) and developed using peroxidase conjugated goat antimo use Ig G and a water insoluble substrate (3,3‐diaminobenzidin tetrahydmchloride; (DAB‐HCI), Nickel chloride and H2O2). the MCIBT gaveL. monocytogenescounts that were not significantly lower than direct colony counts on selective agars. This technique allowed the recovery of 94–100% ofL. monocytogenescells inoculated into foods containing natural background flora counts of 3 × 104to 8 × 106CFU/g or mL. Using a 2 h resuscitation period on nonselective agar before overlay with Oxford media, the MCIBT allowed detection of sublethally heat injured c