Kuo Cheng-Ju, Wang Sin-Tian, Chen Chang-Shi
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University; Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University.
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University; Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University;
J Vis Exp. 2018 Apr 9(134):56169. doi: 10.3791/56169.
Enterohemorrhagic E. coli (EHEC) O157:H7, which is a foodborne pathogen that causesdiarrhea, hemorrhagic colitis (HS), and hemolytic uremic syndrome (HUS), colonize to the intestinal tract of humans. To study the detailed mechanism of EHEC colonization in vivo, it is essential to have animal models to monitor and quantify EHEC colonization. We demonstrate here a mouse-EHEC colonization model by transforming the bioluminescent expressing plasmid to EHEC to monitor and quantify EHEC colonization in living hosts. Animals inoculated with bioluminescence-labeled EHEC show intense bioluminescent signals in mice by detection with a non-invasive in vivo imaging system. After 1 and 2 days post infection, bioluminescent signals could still be detected in infected animals, which suggests that EHEC colonize in hosts for at least 2 days. We also demonstrate that these bioluminescent EHEC locate to mouse intestine, specifically in the cecum and colon, from ex vivo images. This mouse-EHEC colonization model may serve as a tool to advance the current knowledge of the EHEC colonization mechanism.
肠出血性大肠杆菌(EHEC)O157:H7是一种食源性病原体,可导致腹泻、出血性结肠炎(HS)和溶血尿毒综合征(HUS),它会在人类肠道中定殖。为了研究EHEC在体内定殖的详细机制,拥有用于监测和量化EHEC定殖的动物模型至关重要。我们在此展示了一种小鼠-EHEC定殖模型,通过将生物发光表达质粒转化到EHEC中,以监测和量化活体宿主中的EHEC定殖。接种了生物发光标记的EHEC的动物,通过非侵入性体内成像系统检测,在小鼠体内显示出强烈的生物发光信号。感染后1天和2天,在受感染动物中仍可检测到生物发光信号,这表明EHEC在宿主中定殖至少2天。我们还从离体图像中证明,这些生物发光的EHEC定位于小鼠肠道,特别是盲肠和结肠。这种小鼠-EHEC定殖模型可作为一种工具,来推进目前对EHEC定殖机制的认识。