The effect of incorporating accustomed edible mushroom (Agaricus campestris) into processed cheese spread (PCSs) on the chemical, microbiological and organoleptic properties was evaluated. Tiny pieces of mushroom accustomed with steeping into citric acid and boiling in emulsifying salt solution were added to the blend of the cheese spread base at the levels of 5,10 and 15 %. The resulting PCSs were stored at 7°C for 3 months. Significant differences (p < 0.05) were recorded among the chemical composition of PCSs made without and with addition of mushroom. The incorporation of mushroom into PCSs resulted in higher contents of total solids, total protein, SN, ash, total carbohydrates and fiber, as well as pH values than the control spread. On the other hand, control treatment made without mushroom possessed the highest content in F/DM. Addition of mushroom to the base blend did not significantly affect (p > 0.05) in the salt and TVFA contents. The standard plate and psychrotrophic counts of PCSs made without and with mushroom showed slight differences when fresh and during the storage period. The standard plate counts slightly increased during the storage period reaching the maximum counts after one month, and then decreased with prolonged storage. Psychrotrophic bacteria gradually increased in all treatments throughout the storage period. On the other hand, no colonies were found from yeasts and molds, coliform and mesophilic anaerobic spores in all samples examined. Obvious differences (p < 0.05) were noticed in the organoleptic evaluation scores among all treatments of PCSs. The flavours of PCSs with mushroom were generally better and preferable when fresh and throughout the storage period. Addition of 15 % mushroom caused an over pieces of mushroom, which defected the body & texture and appearance & colour of the resulting spread. Therefore, PCSs with improved nutritional and functional values as well as acceptable organoleptic properties and good microbiological quality could be made by incorporation of accustomed edible mushroom into the base blend at the levels of 5 and 10% with refrigerated expiry period more than 3 months.