Bacteria are among the first life forms and ubiquitous on the earth, playing a profound role in maintaining the ecosystem. Among them, pathogenic bacteria cause infectious and fatal bacterial diseases. Zero tolerance limit of some lethal bacterial cells imposed by U.S. regulatory agencies has resulted in the development of numerous detection methods for food and water safety, human health surveillance, and clinical diagnosis of patients. I will talk about a recent approach toward highly sensitive and acurrate counting of pathogenic bacteria using inductively coupled plasma mass spectrometry (ICPMS) in my laboratory, in which alkynyl D-alanine was metabolically assembled into the pathogenic bacterial cell wall via the intrinsic matabolism machinery of bacteria, and an azide-DOTA-europium tag was subsequently click-labeled. Furthermore, corresponding lanthanide-encoded bacterial antibodies were used to recognize the bacteria counted. Five typical bacterial cells of E. coli (ATCC 11775, Escherichia species), S. aureus (ATCC 29213, Staphylococcus species), L. monocytogenes (ATCC 19115, Listeria species), S. dysenteriae (ATCC 13373, Shigella species) and V. parahaemolyticus (ATCC 33847, Vibrio species) were exemplified to demonstrate this proof of concept. Such a strategy developed allows us to directly count dozens to hundreds of targeted pathogenic bacteria using ICPMS.

Metabolic assembly of alkynyl D-alanine into bacterial cell wall and tandem click labeling strategy: Counting pathogenic bacteria using mass spectrometry

256th National Meeting (2018)

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