Enterohemorrhagic Escherichia coli are a dangerous bacterium known to be harmful to the human body, with some infections even resulting in death. Given this danger, food factories are required to perform a quick bacterial test to confirm the absence of this pathogen prior to shipping. We have developed a novel molecular imprinting polymer (MIP) particle that has encapsulated gold nanoparticles (AuNPs) and which can function as both a receptor and optical signal transmitter in biological systems. This MIP particle is artificially synthesized and can be engineered to specifically recognize and capture antigens on the bacterial cell membrane. In addition, MIP particles containing AuNPs generate strong scattered light signals and binding of the MIP particles improves the optical intensity of the target bacterial cells. This enables clear visualization under a darkfield microscope and quantification of the target bacteria using the scattering light intensity. Here we describe the successful quantification of Escherichia coli O157 cells in real meat samples using this technology in conjunction with a simple labelling step.

This paper describes a simple strategy to identify bacteria using the optical properties of the nanohybrid structures (NHs) of polymer-coated metal nanoparticles (NPs). NHs, in which many small NPs are encapsulated in polyaniline particles, are useful optical labels because they produce strong scattered light. The light-scattering characteristics of NHs are strongly dependent on the constituent metal elements of NPs. Gold NHs (AuNHs), silver NHs (AgNHs), and copper NHs (CuNHs) produce white, reddish, and bluish scattered light, respectively. Moreover, unlike NPs, the color of the scattered light does not change even when NHs are aggregated. Introducing an antibody into NHs induces antigen-specific binding to cells, enabling the identification of bacteria based on light scattering. Multiple bacterial species adsorbed on the slide can be identified within a single field of view under a dark field microscope based on the color of the scattered light. Therefore, it is a useful development for safety risk assessments at manufacturing sites, such as those for foods, beverages, and drugs, and environmental surveys that require rapid detection of multiple bacteria.

Gold nanoparticles (AuNPs) are chemically stable and serve as excellent labels because their characteristic red coloration based on the localized surface plasmon resonance does not fade. However, it is necessary...

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A better understanding of the biological functions of microorganism is required to reduce their threats and increase their usefulness. Therefore, an importance of real-time evaluation of bacterial activity increase for various purposes such as hygiene management, development of antibacterial agents, and effective utilization of bacterial resources.1 This necessitates a quantitative assessment of metabolic processes, including growth and respiration. Here we would like to introduce the development of electrochemical methods for assessing bacterial activity. Electrochemical detection of viable bacterial cells was performed using cell membrane permeable electron mediator and redox active pigment. Shewanella oneidensis MR-1 transfers electrons generated within the cell to the extracellular environment via the cytochrome complex in the inner/outer membranes and is one of the most useful bacteria for the recovery of metals, treatment of wastewater, and preparation of microbial fuel cells. By using potentiometric measurements, we have examined intracellular electron generation in bacterial suspensions of S. oneidensis supplemented with different carbon sources or ferricyanide, which was almost completely reduced to ferrocyanide during the incubation without affecting bacterial cell viability.2 On the other hand, a tetrazolium salt (MTT), which was converted to an insoluble reduction form (formazan) through the respiration of microbial cells.3 The insolubility of this formazan was effectively exploited as a surface-confined redox event. The electrochemical detection of formazan was effectively coupled with the thermal lysis of microbes. The sensitivity of the present technique is up to 10,000-fold higher than that of MTT colorimetry and requires an incubation time of only 1 h, which is approximately 1/4 of that required for other metabolism-based techniques. Furthermore, the measurement of the reduction current of dissolved oxygen provides an effective mean for assessing the respiratory activity of bacteria in suspension.4 1) T. Kinoshita, K. Ishiki, D. Q. Nguyen, H. Shiigi, T. Nagaoka, Anal. Chem., 90(6), 4098 (2018). 2) K. Ishiki, H. Shiigi, Anal. Chem., 91(22), 14401-14406 (2019). 3) K. Ishiki, D. Q. Nguyen, A. Morishita, H. Shiigi, T. Nagaoka, Anal. Chem., 90(18), 10903 (2018). 4) M. Saito, K. Ishiki, D. Q. Nguyen, H. Shiigi, Anal. Chem., 91(20), 12793-12798 (2019).