Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis

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-phag...

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Detalles Bibliográficos
Autor principal: Piuri, Mariana
Publicado: 2011
Materias:
enhanced green fluorescent protein
isoniazid
quinolone derivative
rifampicin
streptomycin
accuracy
antibiotic resistance
article
bacterial strain
bacterium detection
bacterium isolate
controlled study
intermethod comparison
mycobacteriophage
Mycobacterium bovis
Mycobacterium smegmatis
Mycobacterium tuberculosis
nonhuman
priority journal
sensitivity analysis
sensitivity and specificity
Antitubercular Agents
Drug Resistance, Bacterial
Fluorometry
Green Fluorescent Proteins
Humans
Microbial Sensitivity Tests
Microbial Viability
Mycobacteriophages
Sensitivity and Specificity
Staining and Labeling
Tuberculosis
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00951137_v49_n5_p1838_Rondon
http://hdl.handle.net/20.500.12110/paper_00951137_v49_n5_p1838_Rondon
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
Descripción
Sumario: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. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

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