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

Piuri, M and Jacobs, WR and Hatfull, GF (2009) Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. PLoS ONE, 4 (3).

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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. © 2009 Piuri et al.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Piuri, M
Jacobs, WR
Hatfull, GFgfh@pitt.eduGFH
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorPai, MadhukarUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Date: 20 March 2009
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 4
Number: 3
DOI or Unique Handle: 10.1371/journal.pone.0004870
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
PubMed Central ID: PMC2654538
PubMed ID: 19300517
Date Deposited: 25 Jul 2012 14:11
Last Modified: 02 Feb 2019 15:58
URI: http://d-scholarship.pitt.edu/id/eprint/13122

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