Real-time video imaging as a new and rapid tool for antibiotic susceptibility testing by the disc diffusion method: a paradigm for evaluating resistance to imipenem and identifying extended-spectrum β-lactamases.

The disc diffusion method has long been considered the standard technique for antibiotic susceptibility testing (AST) in clinical microbiology laboratories because of its simplicity, reproducibility and low cost compared with commercial automated microdilution systems that are usually more rapid but less sensitive for detecting important mechanisms of resistance. Here we measured reading zone diameters around antibiotics in a series of 25 well-characterised Gram-negative bacteria by the disc diffusion technique in real-time using an Advencis Bio-System instrument consisting of a real-time high-resolution video imager in a dedicated incubator. The susceptibility of wild-type Gram-negative bacteria to imipenem, determined by reading the diameter of inhibition, was detectable as early as 3.5h (mean time 3.7 ± 0.45 h), whereas carbapenemase-producing Gram-negative bacteria could be correctly categorised as early as 3h (mean time 4.2 ± 0.8 h) of incubation. Similarly, the characteristic champagne cork aspect of extended-spectrum β-lactamase (ESBL) could be detected by the system as early as 3.5 h. Moreover, we present here for the first time video movies of the appearance of the diameter of inhibition by disc diffusion in real-time. This preliminary study using a new and innovative technology provides for a renewed interest for microbiologists who wish to continue to use the disc diffusion method as a reference method for AST. New video imaging technology presents a proof of concept that could improve the real-time management of patients with AST within a very rapid turnaround time and can provide a large financial saving for hospitals.