The present invention relates to a method for precisely determining protein content in edible fungus, belonging to the technical field of food detection. The method is as follows: determining total nitrogen content in an edible fungus sample by using a Kjeldahl method; additionally taking an equal amount of sample, and washing same with acid and alkali solutions to obtain chitin residues; then determining nitrogen content in the chitin residues by using the Kjeldahl method; subtracting the nitrogen content in the chitin residues from the total nitrogen content in the sample, so as to eliminate the interference of the nitrogen content in chitin; and finally obtaining precise protein content by calculation based on a formula. The present invention provides a method for precisely determining protein content in edible fungus, and provides a method for the precise assessment of the nutrient composition and value of protein in the edible fungus.