A modified direct contact test was used to evaluate the antibacterial properties of four commercially available dental luting agents (RelyX Unicem, Ketac Cem, Ceramir Crown & Bridge and Harvard Cement) and two reference materials (glass-ionomer cement and calcium aluminate cement) compared to a negative-control material (PMMA). Streptococcus mutans bacteria were placed in direct contact with specimens that had been aged for 10 min, 1 day, and 7 days, in order to test the antibacterial properties of the materials. A metabolic assay containing resazurin was used to quantify the amount of viable bacteria remaining after the direct contact tests. The effects of pH and fluoride on bacteria proliferation were also evaluated. Strongest antibacterial properties were found for calcium aluminate cement, followed by Ceramir Crown & Bridge and RelyX Unicem. Ketac Cem, Harvard Cement, and the reference glass-ionomer cement showed bacteria content either higher than or not significantly different from the PMMA control in all instances. pH levels below 6.3 and above 9.0 were found to have negative effects on bacterial proliferation. No correlation between either acidic materials or fluoride release and antibacterial properties could be seen; rather, basic materials showed stronger antibacterial properties.