Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis

Mariana Piuri, William R. Jacobs, Graham F. Hatfull

Research output: Contribution to journalArticle

Abstract

Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis is of paramount importance as multiple- and extensively- drug resistant strains of M. tuberculosis emerge and spread. We describe here a virus-based assay in which fluoromycobacteriophages are used to deliver a GFP or ZsYellow fluorescent marker gene to M. tuberculosis, which can then be monitored by fluorescent detection approaches including fluorescent microscopy and flow cytometry. Pre-clinical evaluations show that addition of either Rifampicin or Streptomycin at the time of phage addition obliterates fluorescence in susceptible cells but not in isogenic resistant bacteria enabling drug sensitivity determination in less than 24 hours. Detection requires no substrate addition, fewer than 100 cells can be identified, and resistant bacteria can be detected within mixed populations. Fluorescence withstands fixation by paraformaldehyde providing enhanced biosafety for testing MDR-TB and XDR-TB infections.

Original languageEnglish (US)
Article numbere4870
JournalPloS one
Volume4
Issue number3
DOIs
StatePublished - Mar 20 2009

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Flow cytometry
Green Fluorescent Proteins
Bacteria
Fluorescence
Anti-Bacterial Agents
Bacteriophages
Testing
Streptomycin
Rifampin
Viruses
Pharmaceutical Preparations
Assays
Microscopic examination
Genes
Substrates
paraform

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. / Piuri, Mariana; Jacobs, William R.; Hatfull, Graham F.

In: PloS one, Vol. 4, No. 3, e4870, 20.03.2009.

Research output: Contribution to journalArticle

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