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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/94323
Ver los metadatos del registro completo
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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 |
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CONICET Digital (CONICET) |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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13.13397 |