Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis

Liliana Rondón, Mariana Piuri, William R. Jacobs, Jacobus De Waard, Graham F. Hatfull, Howard E. Takiff

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We tested a new method for detecting drug-resistant strains of Mycobacterium tuberculosis that uses a TM4 mycobacteriophage phAE87::hsp60-EGFP (EGFP-phage) engineered to contain the gene encoding enhanced green fluorescent protein (EGFP). After promising results in preliminary studies, the EGFP-phage was used to detect isoniazid (INH), rifampin (RIF), and streptomycin (STR) resistance in 155 strains of M. tuberculosis, and the results were compared to the resazurin microplate technique, with the proportion method serving as the reference standard. The resazurin technique yielded sensitivities of 94% for INH and RIF and 98% for STR and specificities of 97% for INH, 95% for RIF, and 98% for STR. The sensitivity of EGFP-phage was 94% for all three antibiotics, with specificities of 90% for INH, 93% for RIF, and 95% for STR. The EGFP-phage results were available in 2 days for RIF and STR and in 3 days for INH, with an estimated cost of ∼2$ to test the three antibiotics. Using a more stringent criterion for resistance improved the specificity of the EGFP-phage for INH and RIF without affecting the sensitivity. In preliminary studies, the EGFP-phage could also effectively detect resistance to the fluoroquinolones. The EGFP-phage method has the potential to be a valuable rapid and economic screen for detecting drug-resistant tuberculosis if the procedure can be simplified, if it can be adapted to clinical material, and if its sensitivity can be improved.

Original languageEnglish (US)
Pages (from-to)1838-1842
Number of pages5
JournalJournal of Clinical Microbiology
Volume49
Issue number5
DOIs
StatePublished - May 1 2011

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Mycobacterium tuberculosis
Drug Resistance
Bacteriophages
Rifampin
Streptomycin
Mycobacteriophages
Anti-Bacterial Agents
enhanced green fluorescent protein
Multidrug-Resistant Tuberculosis
Fluoroquinolones
Isoniazid
Economics
Costs and Cost Analysis
Pharmaceutical Preparations
Genes

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis. / Rondón, Liliana; Piuri, Mariana; Jacobs, William R.; De Waard, Jacobus; Hatfull, Graham F.; Takiff, Howard E.

In: Journal of Clinical Microbiology, Vol. 49, No. 5, 01.05.2011, p. 1838-1842.

Research output: Contribution to journalArticle

Rondón, Liliana ; Piuri, Mariana ; Jacobs, William R. ; De Waard, Jacobus ; Hatfull, Graham F. ; Takiff, Howard E. / Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis. In: Journal of Clinical Microbiology. 2011 ; Vol. 49, No. 5. pp. 1838-1842.
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