Detection and discrimination of <i>Shigella sonnei</i> and <i>Shigella flexneri</i> based on vacuolar responses in <i>Saccharomyces cerevisiae</i>

다국어 초록 (Multilingual Abstract)

<P><B>Abstract</B></P> <P>This study provided a system for bacteria detection based on a lysosome-like-vacuole response in the yeast <I>Saccharomyces cerevisiae</I>. Vacuoles are factors known to activate the immune system in the presence of foreign substances. Here, <I>Shigella sonnei</I> and <I>Shigella flexneri</I> were exposed to yeast to analyze the alteration of vacuolar enzymes. The ability to detect the bacteria was evaluated by confocal microscopy after exposing and staining vacuoles with LysoTracker. Results showed that the treatment of yeast with these bacteria increased the number of red vacuole-like organelles surrounding yeast nuclei. Thus, vacuole alteration can be used as a biomarker for bacteria detection. Next, the expression of vacuolar enzymes under the influence of bacteria was examined using two-dimensional gel electrophoresis (2-DE) method for screening specific biomarkers for each <I>Shigella</I> strain. Finally, the recombinant yeasts that contained biomarkers fused to different fluorescent proteins confirmed the ability of yeast to detect these two <I>Shigella</I> strains at concentrations ranging from 10 to 100 CFU/mL.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Detection of <I>Shigella</I> strains was performed using vacuolar response in <I>S. cerevisiae</I>. </LI> <LI> The vacuolar proteomic in response to <I>Shigella</I> was analyzed for screening of specific biomarkers. </LI> <LI> Biomarker for each strain was fused with different fluorescent proteins for the distinct detection. </LI> <LI> The ability of recombinant yeasts to recognize <I>Shigella</I> strains individually from 10 CFU/ml. </LI> </UL> </P>