Isolation of Mycobacterium avium complex from sputum specimens in association with the appearance of a new cavitary (or infiltrative) lesion was studied in 299 patients from whom the organism was isolated one or more times. Of the patients studied, 114 showed only single isolation. Of these 114, only two patients (2 percent) had association with appearance of a cavitary lesion. Of 29 patients who showed two isolations, 26 (90 percent) had the association. Of 40 patients who showed three isolations, 39 (98 percent) had the association. All 116 patients who showed four or more isolations had the association with appearance of a cavitary lesion. Accordingly, of a total of 185 patients who showed two or more isolations, 181 (98 percent) had the association. Of these 181, 176 (97 percent) showed two or more isolations in the sputum examinations made in the initial three days. Therefore, the sputum examination in the first three days after onset of disease is most important for the diagnosis of disease caused by Mycobacterium avium complex. Since the probability that casual isolation of the organism occurs twice is extremely low, we can make the diagnosis of pulmonary infection caused by this organism by evidence of two or more isolations of the organism in the first few days after the onset of disease, which is associated with appearance of a new cavitary (or infiltrative) lesion. Moreover, theoretical consideration made in this study has led us to conclude that patients who have had a single isolation of the organism together with a new cavitary lesion should be regarded as having an infection.