Pathogenic bacteria can be a major cause of illness from environmental sources as well as the consumption of contaminated products, giving rise to public health concerns globally. The surveillance of such living organisms in food and water supplies remains an important challenge in mitigating their deleterious societal effects. Here, we have developed an optimized bioorthogonal non-canonical amino acid tagging approach to the imaging, capture, and interrogation of shigatoxigenic/verotoxigenicEscherichia coli(VTEC) andListeriathat enables the distinction between living wild-type pathogenic bacteria. The approaches utilize homopropargylglycine (HPG), as well as optimized growth media, that restricts endogenous methionine biosynthesis in a variety of species of public health concern. Endogenous methionine residues are then replaced with HPG, which can then be modified using a myriad of compatible bioorthogonal reactions for tagging of exclusively live bacteria. The methods reported allow for the very rapid screening and identification of living pathogenic organisms.

中文翻译：

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通过优化的生物正交非规范性氨基酸标记快速筛选和鉴定活的病原生物

致病细菌可能是导致环境疾病以及食用受污染产品的主要病因，引起了全球公众健康的关注。对食品和水供应中的这种活生物体的监测仍然是减轻其有害的社会影响的一项重要挑战。在这里，我们已经开发了一种优化的生物正交非规范性氨基酸标签方法，用于对产毒毒素/变毒性大肠杆菌（VTEC）和李斯特菌进行成像，捕获和询问，从而能够区分活的野生型致病细菌。这些方法利用高炔丙基甘氨酸（HPG）以及优化的生长培养基，从而限制了各种公共卫生问题中的内源性蛋氨酸生物合成。然后将内源蛋氨酸残基替换为HPG，然后可以使用无数种兼容的生物正交反应对纯活细菌进行标记，从而对其进行修饰。报告的方法可以非常快速地筛选和鉴定活的病原生物。