The cell-surface protein antigen I/II (Ag I/II) is expressed in oral streptococci, which are known as the causative agent of a number of diseases including dental caries, endocarditis, gingivitis, and periodontal disease. Consequently, monoclonal antibodies (MAb) capable of recognizing the streptococcal Ag I/II protein could be a useful tool for the diagnosis and cure of these diseases. In this study, a previously generated monoclonal anti-Ag I/II antibody, ckAg I/II, was used to detect a small amount of Streptococcus mutans (S. mutans) surface antigen Ag I/II. The ckAg I/II was proved to be very sensitive and able to detect as little as 1 ng of recombinant Ag I/II protein within 5 min and Ag I/II in saliva within 10 min, as well as native Ag I/II in 20 μL of culture supernatant by ELISA. These results suggest that ckAg I/II can be used as a fast and efficient diagnostic tool to detect Ag I/II.