φ 2GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples

Paras Jain, Travis E. Hartman, Nell Eisenberg, Max R. O'Donnell, Jordan Kriakov, Karnishree Govender, Mantha Makume, David S. Thaler, Graham F. Hatfull, A. Willem Sturm, Michelle H. Larsen, Preshnie Moodley, William R. Jacobs

Research output: Contribution to journalArticle

47 Scopus citations

Abstract

The difficulty of diagnosing active tuberculosis (TB) and lack of rapid drug susceptibility testing (DST) at the point of care remain critical obstacles to TB control. This report describes a high-intensity mycobacterium-specific- fluorophage (φ 2GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing φ 2GFP10 from previous reporter phages include an improved vector backbone with increased cloning capacity and superior expression of fluorescent reporter genes through use of an efficient phage promoter. φ 2GFP10 produces a 100-fold increase in fluorescence per cell compared to existing reporter phages. DST for isoniazid and oxofloxacin, carried out in cultured samples, was complete within 36 h. Use of φ 2GFP10 detected M. tuberculosis in clinical sputum samples collected from TB patients. DST for rifampin and kanamycin from sputum samples yielded results after 12 h of incubation with φ 2GFP10. Fluorophage φ 2GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection and for rapid DST.

Original languageEnglish (US)
Pages (from-to)1362-1369
Number of pages8
JournalJournal of Clinical Microbiology
Volume50
Issue number4
DOIs
StatePublished - Apr 2012

ASJC Scopus subject areas

  • Microbiology (medical)

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    Jain, P., Hartman, T. E., Eisenberg, N., O'Donnell, M. R., Kriakov, J., Govender, K., Makume, M., Thaler, D. S., Hatfull, G. F., Sturm, A. W., Larsen, M. H., Moodley, P., & Jacobs, W. R. (2012). φ 2GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples. Journal of Clinical Microbiology, 50(4), 1362-1369. https://doi.org/10.1128/JCM.06192-11