2. Academic Validation
  3. Rapid and specific labeling of single live Mycobacterium tuberculosis with a dual-targeting fluorogenic probe

Rapid and specific labeling of single live Mycobacterium tuberculosis with a dual-targeting fluorogenic probe

  • Sci Transl Med. 2018 Aug 15;10(454):eaar4470. doi: 10.1126/scitranslmed.aar4470.
Yunfeng Cheng 1,  Jinghang Xie 1,  Kyung-Hyun Lee 1 2,  Rajiv L Gaur 3 4 5,  Aiguo Song 1,  Tingting Dai 1,  Hongjun Ren 6,  Jiannan Wu 7 8,  Zhaogang Sun 7 8,  Niaz Banaei 3 4 5,  Demir Akin 9,  Jianghong Rao 10
Affiliations

Affiliations

  • 1 Departments of Radiology and Chemistry, Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 2 Institute of Bioengineering and Nanotechnology, The Nanos, Singapore 138669, Singapore.
  • 3 Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 4 Clinical Microbiology Laboratory, Stanford University Medical Center, Palo Alto, CA 94304, USA.
  • 5 Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 6 Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou 310018, P. R. China.
  • 7 National Tuberculosis Clinical Laboratory, Beijing Chest Hospital, Capital Medical University, Beijing 101149, P. R. China.
  • 8 Beijing Key Laboratory for Drug Resistance Tuberculosis Research, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing 101149, P. R. China.
  • 9 Center for Cancer Nanotechnology Excellence, Department of Radiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
  • 10 Departments of Radiology and Chemistry, Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, CA 94305, USA. [email protected].
PMID: 30111644 DOI: 10.1126/scitranslmed.aar4470
Abstract

Tuberculosis (TB) remains a public health crisis and a leading cause of infection-related death globally. Although in high demand, imaging technologies that enable rapid, specific, and nongenetic labeling of live Mycobacterium tuberculosis (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.

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