Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis

Autores
Rondon Salazar, Liliana; Piuri, Mariana; Jacobs Jr., William R.; De Waard, Jacobus; Hatfull, Graham F.; Takiff, Howard Eugene
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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.
Fil: Rondon Salazar, Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Piuri, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Jacobs Jr., William R.. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: De Waard, Jacobus. Hospital Vargas; Venezuela
Fil: Hatfull, Graham F.. University of Pittsburgh; Estados Unidos
Fil: Takiff, Howard Eugene. Instituto Venezolano de Investigaciones Científicas; Venezuela
Materia
Mycobacteriophages
Mycobacterium tuberculosis
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/94323

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network_name_str CONICET Digital (CONICET)
spelling Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosisRondon Salazar, LilianaPiuri, MarianaJacobs Jr., William R.De Waard, JacobusHatfull, Graham F.Takiff, Howard EugeneMycobacteriophagesMycobacterium tuberculosishttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We 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.Fil: Rondon Salazar, Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; VenezuelaFil: Piuri, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; VenezuelaFil: Jacobs Jr., William R.. Instituto Venezolano de Investigaciones Científicas; VenezuelaFil: De Waard, Jacobus. Hospital Vargas; VenezuelaFil: Hatfull, Graham F.. University of Pittsburgh; Estados UnidosFil: Takiff, Howard Eugene. Instituto Venezolano de Investigaciones Científicas; VenezuelaAmerican Society for Microbiology2011-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/94323Rondon Salazar, Liliana; Piuri, Mariana; Jacobs Jr., William R.; De Waard, Jacobus; Hatfull, Graham F.; et al.; Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis; American Society for Microbiology; Journal of Clinical Microbiology; 49; 5; 5-2011; 1838-18420095-1137CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://jcm.asm.org/content/49/5/1838info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.02476-10info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:58Zoai:ri.conicet.gov.ar:11336/94323instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:02:59.032CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
title Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
spellingShingle Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis
Rondon Salazar, Liliana
Mycobacteriophages
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
dc.creator.none.fl_str_mv Rondon Salazar, Liliana
Piuri, Mariana
Jacobs Jr., William R.
De Waard, Jacobus
Hatfull, Graham F.
Takiff, Howard Eugene
author Rondon Salazar, Liliana
author_facet Rondon Salazar, Liliana
Piuri, Mariana
Jacobs Jr., William R.
De Waard, Jacobus
Hatfull, Graham F.
Takiff, Howard Eugene
author_role author
author2 Piuri, Mariana
Jacobs Jr., William R.
De Waard, Jacobus
Hatfull, Graham F.
Takiff, Howard Eugene
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Mycobacteriophages
Mycobacterium tuberculosis
topic Mycobacteriophages
Mycobacterium tuberculosis
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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.
Fil: Rondon Salazar, Liliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Piuri, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: Jacobs Jr., William R.. Instituto Venezolano de Investigaciones Científicas; Venezuela
Fil: De Waard, Jacobus. Hospital Vargas; Venezuela
Fil: Hatfull, Graham F.. University of Pittsburgh; Estados Unidos
Fil: Takiff, Howard Eugene. Instituto Venezolano de Investigaciones Científicas; Venezuela
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.
publishDate 2011
dc.date.none.fl_str_mv 2011-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/94323
Rondon Salazar, Liliana; Piuri, Mariana; Jacobs Jr., William R.; De Waard, Jacobus; Hatfull, Graham F.; et al.; Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis; American Society for Microbiology; Journal of Clinical Microbiology; 49; 5; 5-2011; 1838-1842
0095-1137
CONICET Digital
CONICET
url http://hdl.handle.net/11336/94323
identifier_str_mv Rondon Salazar, Liliana; Piuri, Mariana; Jacobs Jr., William R.; De Waard, Jacobus; Hatfull, Graham F.; et al.; Evaluation of fluoromycobacteriophages for detecting drug resistance in Mycobacterium tuberculosis; American Society for Microbiology; Journal of Clinical Microbiology; 49; 5; 5-2011; 1838-1842
0095-1137
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://jcm.asm.org/content/49/5/1838
info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.02476-10
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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