Science Translational Medicine · 2018 · 88 citations · 29 references
Natural SciencesCell BiologyMicrobiologyDiagnostic MicrobiologyMolecular ImagingSingle-molecule BiophysicsFluorescence ImagingFluorescence MicroscopyMicrofluidic BiosensorsMicrofluidicsSingle-molecule DetectionSingle-molecule Fluorescence MicroscopyFluorescent Cell LabelingLive-cell Fluorescence ImagingLive-cell Imaging ProbesLive-cell LabelingMicrofluidic AnalysisMicrofluidic Chip DesignMicrofluidic PlatformsMicrofluidic Single-cell Analysis
(Mtb) remain underdeveloped. We report a dual-targeting strategy to develop a small molecular probe (CDG-DNB3) that can fluorescently label single bacilli within 1 hour. CDG-DNB3 fluoresces upon activation of the β-lactamase BlaC, a hydrolase naturally expressed in Mtb, and the fluorescent product is retained through covalent modification of the Mtb essential enzyme decaprenylphosphoryl-β-d-ribose 2'-epimerase (DprE1). This dual-targeting probe not only discriminates live from dead Bacillus Calmette-Guérin (BCG) but also shows specificity for Mtb over other bacterial species including 43 nontuberculosis mycobacteria (NTM). In addition, CDG-DNB3 can image BCG phagocytosis in real time, as well as Mtb in patients' sputum. Together with a low-cost, self-driven microfluidic chip, we have achieved rapid labeling and automated quantification of live BCG. This labeling approach should find many potential applications for research toward TB pathogenesis, treatment efficacy assessment, and diagnosis.
29
Benzothiazinones Kill <i>Mycobacterium tuberculosis</i> by Blocking Arabinan Synthesis
Vadim Makarov, Giulia Manina, Katarı́na Mikušová et al. · Science · 2009 · 727 citations · Full text