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Sci Transl Med. 2018 Feb 28;10(430). pii: eaam6310. doi: 10.1126/scitranslmed.aam6310.

Rapid detection of Mycobacterium tuberculosis in sputum with a solvatochromic trehalose probe.

Author information

1
Department of Biology, Stanford University, Stanford, CA 94305, USA.
2
Department of Chemistry, Stanford University, Stanford, CA 94305, USA.
3
Department of Science and Technology/National Research Foundation Centre of Excellence for Biomedical Tuberculosis Research, Faculty of Health Sciences, University of Witwatersrand, National Health Laboratory Service, Johannesburg, South Africa.
4
Institute of Pharmaceutical Biology and Biotechnology, Heinrich Heine University Duesseldorf, Universitätsstrasse 1, 40225 Duesseldorf, Germany.
5
Department of Microbiology and Immunology, Stanford University, Stanford, CA 94305, USA.
6
Perinatal HIV Research Unit (PHRU), SA MRC Soweto Matlosana Collaborating Centre for HIV/AIDS and TB, University of the Witwatersrand, Johannesburg, South Africa.
7
Medical Research Council-Centre for the AIDS Programme of Research in South Africa (CAPRISA) HIV-TB Pathogenesis and Treatment Research Unit, Durban, South Africa.
8
Department of Chemistry, Stanford University, Stanford, CA 94305, USA. bertozzi@stanford.edu.
9
Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.

Abstract

Tuberculosis (TB) is the leading cause of death from an infectious bacterial disease. Poor diagnostic tools to detect active disease plague TB control programs and affect patient care. Accurate detection of live Mycobacterium tuberculosis (Mtb), the causative agent of TB, could improve TB diagnosis and patient treatment. We report that mycobacteria and other corynebacteria can be specifically detected with a fluorogenic trehalose analog. We designed a 4-N,N-dimethylamino-1,8-naphthalimide-conjugated trehalose (DMN-Tre) probe that undergoes >700-fold increase in fluorescence intensity when transitioned from aqueous to hydrophobic environments. This enhancement occurs upon metabolic conversion of DMN-Tre to trehalose monomycolate and incorporation into the mycomembrane of Actinobacteria. DMN-Tre labeling enabled the rapid, no-wash visualization of mycobacterial and corynebacterial species without nonspecific labeling of Gram-positive or Gram-negative bacteria. DMN-Tre labeling was detected within minutes and was inhibited by heat killing of mycobacteria. Furthermore, DMN-Tre labeling was reduced by treatment with TB drugs, unlike the clinically used auramine stain. Lastly, DMN-Tre labeled Mtb in TB-positive human sputum samples comparably to auramine staining, suggesting that this operationally simple method may be deployable for TB diagnosis.

PMID:
29491187
PMCID:
PMC5985656
DOI:
10.1126/scitranslmed.aam6310
[Indexed for MEDLINE]
Free PMC Article

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