Electrochemical Evaluation of the Number of Viable Bacteria Using Carbon Electrode Chip

Abstract

To perform on-site bacterial testing at food and pharmaceutical manufacturing sites, it is desired to develop a new method that can quickly measure the number of viable bacterial cells. We have succeeded in measuring the number of viable bacteria using small and inexpensive disposable electrode chips focusing on electrochemical methods that realize quick detection and device miniaturization. The oxidized form of the tetrazolium salt, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), is soluble and highly permeable through cell membranes. MTT is a useful indicator for evaluating cell activity that not only turns color as a result of structural changes related to intracellular metabolism but also causes a clear current response. The carbon-screen-printed electrode chip provides a distinct current response related to the MTT redox reaction in a small volume of bacterial suspension (50 µL). Based on the fact that the current reaction of MTT was strongly dependent on intracellular metabolism, the number of viable cells in a bacterial suspension could be measured electrochemically. Current changes for live cells occurred within 10 min and increased with the incubation time. After only 60 min of incubation, we successfully estimated the number of viable cells in a bacterial suspension of 10 3 CFU mL −1 . This technology eliminates the need for complicated testing, expensive equipment, and lengthy culture testing times, thereby enabling the confirmation of food safety before shipping to prevent food poisoning.