Toward Point-of-Care Detection of Mycobacterium tuberculosis: A Brighter Solvatochromic Probe Detects Mycobacteria within Minutes

Mireille Kamariza; Samantha G L Keyser; Ashley Utz; Benjamin D Knapp; Christopher Ealand; Green Ahn; C J Cambier; Teresia Chen; Bavesh Kana; Kerwyn Casey Huang; Carolyn R Bertozzi

doi:10.1021/jacsau.1c00173

Toward Point-of-Care Detection of Mycobacterium tuberculosis: A Brighter Solvatochromic Probe Detects Mycobacteria within Minutes

JACS Au. 2021 Jul 26;1(9):1368-1379. doi: 10.1021/jacsau.1c00173. eCollection 2021 Sep 27.

Authors

Mireille Kamariza¹, Samantha G L Keyser², Ashley Utz¹, Benjamin D Knapp³, Christopher Ealand⁴, Green Ahn⁵, C J Cambier⁵, Teresia Chen¹, Bavesh Kana^{4

6}, Kerwyn Casey Huang^{3

7

8

9}, Carolyn R Bertozzi^{2

5

10}

Affiliations

¹ Department of Biology, Stanford University, Stanford, California 94305, United States.
² Department of Chemistry, University of California, Berkeley, Berkeley, California 94720, United States.
³ Biophysics Program, Stanford University, Stanford, California 94305, United States.
⁴ 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 2000, South Africa.
⁵ Department of Chemistry, Stanford University, Stanford, California 94305, United States.
⁶ Medical Research Council-Centre for the AIDS Programme of Research in South Africa (CAPRISA) HIV-TB Pathogenesis and Treatment Research Unit, Durban 4013, South Africa.
⁷ Department of Bioengineering, Stanford University, Stanford, California 94305, United States.
⁸ Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305, United States.
⁹ Chan Zuckerberg Biohub, San Francisco, California 94158, United States.
¹⁰ Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, United States.

Abstract

There is an urgent need for point-of-care tuberculosis (TB) diagnostic methods that are fast, inexpensive, and operationally simple. Here, we report on a bright solvatochromic dye trehalose conjugate that specifically detects Mycobacterium tuberculosis (Mtb) in minutes. 3-Hydroxychromone (3HC) dyes, known for having high fluorescence quantum yields, exhibit shifts in fluorescence intensity in response to changes in environmental polarity. We synthesized two analogs of 3HC-trehalose conjugates (3HC-2-Tre and 3HC-3-Tre) and determined that 3HC-3-Tre has exceptionally favorable properties for Mtb detection. 3HC-3-Tre-labeled mycobacterial cells displayed a 10-fold increase in fluorescence intensity compared to our previous reports on the dye 4,4-N,N-dimethylaminonapthalimide (DMN-Tre). Excitingly, we detected fluorescent Mtb cells within 10 min of probe treatment. Thus, 3HC-3-Tre permits rapid visualization of mycobacteria that ultimately could empower improved Mtb detection at the point-of-care in low-resource settings.

Grants and funding

R37 AI051622/AI/NIAID NIH HHS/United States