Real-time monitoring of bacterial growth and fast antimicrobial susceptibility tests exploiting multiple light scattering

Abstract: Antimicrobial susceptibility test (AST) is widely used to provide the minimum inhibitory concentration of bacteria, and crucial to provide appropriate uses of antibiotics and to address the issue of drug-resistance bacteria. However, ASTs require the time-consuming incubation about 16-20 h for the visual determination of the growth of bacterial colonies, which has been a major obstacle to on-site applications of ASTs. In this study, we propose a rapid and non-invasive method based on laser speckles to evaluate the bacterial growth movements in real time, thus reducing the time for the agar dilution method. With a simple configuration compatible with conventional agar plates, the analysis of laser speckle from samples enables the early detection of the presence of growth as well as its detailed history of the colony-forming movement on agar plates. Using the samples prepared through the same procedure as the agar dilution method, we obtained the AST results at least 4-8 hours earlier than the conventional method without compromising the accuracy. This technique does not require for the use of exogenous agents, but works for most bacteria regardless of their species. Furthermore, the distinctive responses of several species to microbial agents were revealed through the present technique supporting a comprehensive analysis of the effect of the antibiotics. The findings suggest that this new method could be a useful tool for rapid, simple, and low-cost ASTs in addition to providing the historical information of the bacterial growth on agar plates.