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spelling paper:paper_00951137_v49_n5_p1838_Rondon2023-06-08T15:09:38Z Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis Piuri, Mariana 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 Mycobacterium tuberculosis Sensitivity and Specificity Staining and Labeling Tuberculosis 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-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. Fil:Piuri, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2011 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
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic 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
Mycobacterium tuberculosis
Sensitivity and Specificity
Staining and Labeling
Tuberculosis
Mycobacterium tuberculosis
spellingShingle 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
Mycobacterium tuberculosis
Sensitivity and Specificity
Staining and Labeling
Tuberculosis
Mycobacterium tuberculosis
Piuri, Mariana
Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
topic_facet 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
Mycobacterium tuberculosis
Sensitivity and Specificity
Staining and Labeling
Tuberculosis
Mycobacterium tuberculosis
description 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.
author Piuri, Mariana
author_facet Piuri, Mariana
author_sort Piuri, Mariana
title Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title_short Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title_full Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title_fullStr Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title_full_unstemmed Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title_sort evaluation of fluoromycobacteriophages for detecting drug resistance in mycobacterium tuberculosis
publishDate 2011
url 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
work_keys_str_mv AT piurimariana evaluationoffluoromycobacteriophagesfordetectingdrugresistanceinmycobacteriumtuberculosis
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