The aim of this study was to evaluate the antibacterial action of root canal mechanical preparation using 2.5% sodium hypochlorite as the irrigating solution and a calcium hydroxide paste as the antibacterial intracanal dressing in human primary teeth root canals with pulp necrosis and apical periodontitis by means of microbial culture. A total of 26 root canals of human primary teeth with pulp necrosis and apical periodontitis were used. Samples were collected before, 72h after biomechanical treatment and 72h after removal of the intracanal dressing. Comparison by Wilcoxon test showed that root canal mechanical preparation effectively eliminated all microorganisms in 20% of the root canals, and the intracanal dressing in 62.5%; however, the cumulative action of biomechanical treatment and intracanal dressing eliminated the microorganisms of 70% of the root canals (p<0.001). Isolated root canal mechanical preparation presented poorer microbiological results that those obtained with root canal mechanical preparation and the use of an intracanal dressing indicating the necessity of topical application of an intracanal medication between sessions in primary teeth with pulp necrosis and apical periodontitis.