SETTING: Patients with suspected pulmonary tuberculosis (TB) visiting government TB diagnostic and treatment centres in Makassar City, South Sulawesi Province, Indonesia, from February to October 2008 were included in the study. OBJECTIVE: To determine the distribution of Mycobacterium tuberculosis genotypes in Makassar. DESIGN: Cross-sectional study. Spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) and principal genetic grouping (PGG) were used to genotype the M. tuberculosis clinical isolates. RESULTS: Among 179 isolates derived from pulmonary TB patients, distribution of spoligotypes comprised the East Africa-Indian (30.2%), T (17.9%), H (12.3%) and Beijing (9.5%) lineages. Other lineages found in smaller proportions were the Latin American-Mediterranean, MANU, S and X lineages. Nineteen isolates (10.6%) could not be grouped into any of the reported lineages or shared types. Single nucleotide polymorphism analysis of katG(463) and gyrA(95) grouped these isolates primarily into PGG1 (9/19, 47%). CONCLUSION: Only a few genetically identical clustered isolates were identified within the 9-month study period, and most isolates were genetically diverse. Furthermore, 15 spoligopatterns identified in our study have not been reported previously. To our knowledge, this is the first comprehensive study describing genotypes of M. tuberculosis clinical isolates in Sulawesi.