Real-time imaging of mycobacterium tuberculosis, using a novel near-infrared fluorescent substrate

Hee Jeong Yang, Ying Kong, Yunfeng Cheng, Harish Janagama, Hany Hassounah, Hexin Xie, Jianghong Rao, Jeffrey D. Cirillo

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

California Slow growth of Mycobacterium tuberculosis, the causative agent of tuberculosis, hinders advancement in all areas of research toward prevention and treatment. Real-time imaging with reporter enzyme fluorescence (REF) that uses custom fluorogenic substrates for bacterial enzymes allows rapid and specific detection of M. tuberculosis in live animals. We have synthesized a novel REF substrate, CNIR800, that carries a near-infrared (NIR) fluorochrome IRDye 800CW, with a quencher connected through the lactam ring that is hydrolyzed by the enzyme BlaC (β-lactamase) that is naturally expressed by M. tuberculosis. CNIR800 produces long-wavelength emission at 795 nm upon excitation (745 nm) and exhibits significantly improved signal to noise ratios for detection of M. tuberculosis. The detection threshold with CNIR800 is approximately 100 colony-forming units (CFU) in vitro and <1000 CFU in the lungs of mice. Additionally, fluorescence signal from cleaved CNIR800 reaches maximal levels 4-6 hours after administration in live animals, allowing accurate evaluation of antituberculous drug efficacy. Thus, CNIR800 represents an excellent substrate for accurate detection of M. tuberculosis rapidly and specifically in animals, facilitating research toward understanding pathogenic mechanisms, evaluation of therapeutic outcomes, and screening new vaccines.

Original languageEnglish (US)
Pages (from-to)405-414
Number of pages10
JournalJournal of Infectious Diseases
Volume215
Issue number3
DOIs
StatePublished - Jan 1 2017

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Mycobacterium tuberculosis
Fluorescence
Enzymes
Fluorescent Dyes
Stem Cells
Lactams
Drug Evaluation
Signal-To-Noise Ratio
Tuberculosis
Vaccines
Lung
Growth
Research
Therapeutics

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Infectious Diseases

Cite this

Real-time imaging of mycobacterium tuberculosis, using a novel near-infrared fluorescent substrate. / Yang, Hee Jeong; Kong, Ying; Cheng, Yunfeng; Janagama, Harish; Hassounah, Hany; Xie, Hexin; Rao, Jianghong; Cirillo, Jeffrey D.

In: Journal of Infectious Diseases, Vol. 215, No. 3, 01.01.2017, p. 405-414.

Research output: Contribution to journalArticle

Yang, HJ, Kong, Y, Cheng, Y, Janagama, H, Hassounah, H, Xie, H, Rao, J & Cirillo, JD 2017, 'Real-time imaging of mycobacterium tuberculosis, using a novel near-infrared fluorescent substrate', Journal of Infectious Diseases, vol. 215, no. 3, pp. 405-414. https://doi.org/10.1093/infdis/jiw298
Yang, Hee Jeong ; Kong, Ying ; Cheng, Yunfeng ; Janagama, Harish ; Hassounah, Hany ; Xie, Hexin ; Rao, Jianghong ; Cirillo, Jeffrey D. / Real-time imaging of mycobacterium tuberculosis, using a novel near-infrared fluorescent substrate. In: Journal of Infectious Diseases. 2017 ; Vol. 215, No. 3. pp. 405-414.
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