Background: Mycobacterium africanum continue to cause many cases of tuberculosis in Central West African countries, while in Cameroon, it’s decreasing and M. tuberculosis mostly represented by a “Cameroon family” is increasing. Actually these observations were based in studies done in few regions of Cameroon. Materials and findings: A cross section study was therefore done in Douala one of the most cosmopolite town of Cameroon. 707 patients consulting for tuberculosis were examined and sputum was collected. Acid-fast bacilli (AFB) were detected on Ziehl-Neelsen stained impression smear and culture was made in Löwenstein-Jensen and in Löwenstein-Jensen media supplemented with 0.4% of pyruvate. Isolated strains were typed using spoligotyping and MIRU-VNTR typing. Among the 707 sputa collected, 371 were positive in culture. The molecular typing revealed that 97.3% (354/364) of cases were due to M. tuberculosis mostly represented by ST61 clad and 2.74% (10/364) to M. africanum. No M. bovis was present. Conclusion: This trend shows a high proportion of Cameroon family and evidence the low contribution of M. africanum on pulmonary TB compared to others region of Cameroon.