Objective To revive the criteria for neostigmine test,and prove a scientific,accurate,applicable method to help diagnosis of myasthenia gravis(MG).Methods Neostigmine(1.5 mg) and atropine(1.0 mg) were intramuscularly administered for 68 cases with clinical confirmed MG and 23 healthy control cases.The clinical absolute and relative scores of each item were recorded quantitatively before and every 10 minutes after intramuscular injection of neostigmine.Results The test result was positive when each item relative score was over 60%,25%-60% was possible positive and under 25% was negative.Conclusions The criteria of neostigmine test established in this study is objective,accurate,easy and applicable for MG diagnosis.