Comparative study of rapid ATP bioluminescence assay and conventional plate count method for development of rapid disinfecting activity test.

Abstract

Antimicrobial activity is tested when developing disinfectants, pharmaceutical products, cosmetics, and many other consumer products. However, the plate count method, the conventional way to count the number of microorganisms, needs several days of culture. Consequently, a means of rapid microbial detection is strongly desired to replace this method. We have already developed a rapid and sensitive microbial adenosine triphosphate (ATP) detection system utilizing ATP bioluminescence, which can quantify microbial ATP within 1 h. To apply this technique to antibacterial activity tests, the ATP method should be proved equal or superior to the conventional method. In this study, we conducted disinfectant activity tests comparing the ATP method and the plate count method, using polyhexamethylene biguanide (PHMB) in different concentrations (0-10 ppm) as a model disinfectant against Staphylococcus aureus and Aspergillus brasiliensis. We found that the log reduction of intracellular ATP had a positive correlation with the log reduction of the plate count. Moreover, the ATP method was able to distinguish different conditions of injured microbial cells that were observed using scanning electron microscopy, whereas colony counting detects only culturable cells. The ATP method is thus a rapid and useful alternative to the conventional method in the field of antimicrobial activity testing.