Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing

Autores
Engström, Anna; Morcillo, Nora Susana; Imperiale, Belén Rocío; Hoffner, Sven E.; Juréen, Pontus
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Conventional phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and very time-consuming. Early detection of drug-resistant tuberculosis (TB) is essential for prevention and control of TB transmission. We have developed a pyrosequencing method for simultaneous detection of mutations associated with resistance to rifampin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and ofloxacin. Seven pyrosequencing assays were optimized for following loci: rpoB, katG, embB, rrs, gyrA, and the promoter regions of inhA and eis. The molecular method was evaluated on a panel of 290 clinical isolates of M. tuberculosis. In comparison to phenotypic DST, the pyrosequencing method demonstrated high specificity (100%) and sensitivity (94.6%) for detection of multidrug-resistant M. tuberculosis as well as high specificity (99.3%) and sensitivity (86.9%) for detection of extensively drug-resistant M. tuberculosis. The short turnaround time combined with multilocus sequencing of several isolates in parallel makes pyrosequencing an attractive method for drug resistance screening in M. tuberculosis.
Fil: Engström, Anna. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
Fil: Morcillo, Nora Susana. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; Argentina
Fil: Imperiale, Belén Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; Argentina
Fil: Hoffner, Sven E.. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
Fil: Juréen, Pontus. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
Materia
MYCOBACTERIUM TUBERCULOSIS
PYROSEQUENCING
DRUG RESISTANCE
FIRST AND SECOND LINE DRUGS
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:11336/192466
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencingEngström, AnnaMorcillo, Nora SusanaImperiale, Belén RocíoHoffner, Sven E.Juréen, PontusMYCOBACTERIUM TUBERCULOSISPYROSEQUENCINGDRUG RESISTANCEFIRST AND SECOND LINE DRUGShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Conventional phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and very time-consuming. Early detection of drug-resistant tuberculosis (TB) is essential for prevention and control of TB transmission. We have developed a pyrosequencing method for simultaneous detection of mutations associated with resistance to rifampin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and ofloxacin. Seven pyrosequencing assays were optimized for following loci: rpoB, katG, embB, rrs, gyrA, and the promoter regions of inhA and eis. The molecular method was evaluated on a panel of 290 clinical isolates of M. tuberculosis. In comparison to phenotypic DST, the pyrosequencing method demonstrated high specificity (100%) and sensitivity (94.6%) for detection of multidrug-resistant M. tuberculosis as well as high specificity (99.3%) and sensitivity (86.9%) for detection of extensively drug-resistant M. tuberculosis. The short turnaround time combined with multilocus sequencing of several isolates in parallel makes pyrosequencing an attractive method for drug resistance screening in M. tuberculosis.Fil: Engström, Anna. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; SueciaFil: Morcillo, Nora Susana. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; ArgentinaFil: Imperiale, Belén Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; ArgentinaFil: Hoffner, Sven E.. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; SueciaFil: Juréen, Pontus. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; SueciaAmerican Society for Microbiology2012-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/192466Engström, Anna; Morcillo, Nora Susana; Imperiale, Belén Rocío; Hoffner, Sven E.; Juréen, Pontus; Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing; American Society for Microbiology; Journal of Clinical Microbiology; 50; 6; 6-2012; 2026-20330095-1137CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://journals.asm.org/doi/10.1128/JCM.06664-11info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.06664-11info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:44:16Zoai:ri.conicet.gov.ar:11336/192466instacron: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:34982025-09-29 09:44:16.781CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
title Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
spellingShingle Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
Engström, Anna
MYCOBACTERIUM TUBERCULOSIS
PYROSEQUENCING
DRUG RESISTANCE
FIRST AND SECOND LINE DRUGS
title_short Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
title_full Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
title_fullStr Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
title_full_unstemmed Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
title_sort Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing
dc.creator.none.fl_str_mv Engström, Anna
Morcillo, Nora Susana
Imperiale, Belén Rocío
Hoffner, Sven E.
Juréen, Pontus
author Engström, Anna
author_facet Engström, Anna
Morcillo, Nora Susana
Imperiale, Belén Rocío
Hoffner, Sven E.
Juréen, Pontus
author_role author
author2 Morcillo, Nora Susana
Imperiale, Belén Rocío
Hoffner, Sven E.
Juréen, Pontus
author2_role author
author
author
author
dc.subject.none.fl_str_mv MYCOBACTERIUM TUBERCULOSIS
PYROSEQUENCING
DRUG RESISTANCE
FIRST AND SECOND LINE DRUGS
topic MYCOBACTERIUM TUBERCULOSIS
PYROSEQUENCING
DRUG RESISTANCE
FIRST AND SECOND LINE DRUGS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Conventional phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and very time-consuming. Early detection of drug-resistant tuberculosis (TB) is essential for prevention and control of TB transmission. We have developed a pyrosequencing method for simultaneous detection of mutations associated with resistance to rifampin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and ofloxacin. Seven pyrosequencing assays were optimized for following loci: rpoB, katG, embB, rrs, gyrA, and the promoter regions of inhA and eis. The molecular method was evaluated on a panel of 290 clinical isolates of M. tuberculosis. In comparison to phenotypic DST, the pyrosequencing method demonstrated high specificity (100%) and sensitivity (94.6%) for detection of multidrug-resistant M. tuberculosis as well as high specificity (99.3%) and sensitivity (86.9%) for detection of extensively drug-resistant M. tuberculosis. The short turnaround time combined with multilocus sequencing of several isolates in parallel makes pyrosequencing an attractive method for drug resistance screening in M. tuberculosis.
Fil: Engström, Anna. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
Fil: Morcillo, Nora Susana. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; Argentina
Fil: Imperiale, Belén Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Ministerio de Salud. Hospital "Dr. Antonio A. Cetrángolo"; Argentina
Fil: Hoffner, Sven E.. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
Fil: Juréen, Pontus. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia. Swedish Institute for Communicable Disease Control; Suecia
description Conventional phenotypic drug susceptibility testing (DST) methods for Mycobacterium tuberculosis are laborious and very time-consuming. Early detection of drug-resistant tuberculosis (TB) is essential for prevention and control of TB transmission. We have developed a pyrosequencing method for simultaneous detection of mutations associated with resistance to rifampin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and ofloxacin. Seven pyrosequencing assays were optimized for following loci: rpoB, katG, embB, rrs, gyrA, and the promoter regions of inhA and eis. The molecular method was evaluated on a panel of 290 clinical isolates of M. tuberculosis. In comparison to phenotypic DST, the pyrosequencing method demonstrated high specificity (100%) and sensitivity (94.6%) for detection of multidrug-resistant M. tuberculosis as well as high specificity (99.3%) and sensitivity (86.9%) for detection of extensively drug-resistant M. tuberculosis. The short turnaround time combined with multilocus sequencing of several isolates in parallel makes pyrosequencing an attractive method for drug resistance screening in M. tuberculosis.
publishDate 2012
dc.date.none.fl_str_mv 2012-06
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/11336/192466
Engström, Anna; Morcillo, Nora Susana; Imperiale, Belén Rocío; Hoffner, Sven E.; Juréen, Pontus; Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing; American Society for Microbiology; Journal of Clinical Microbiology; 50; 6; 6-2012; 2026-2033
0095-1137
CONICET Digital
CONICET
url http://hdl.handle.net/11336/192466
identifier_str_mv Engström, Anna; Morcillo, Nora Susana; Imperiale, Belén Rocío; Hoffner, Sven E.; Juréen, Pontus; Detection of first- and second-line drug resistance in Mycobacterium tuberculosis clinical isolates by pyrosequencing; American Society for Microbiology; Journal of Clinical Microbiology; 50; 6; 6-2012; 2026-2033
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://journals.asm.org/doi/10.1128/JCM.06664-11
info:eu-repo/semantics/altIdentifier/doi/10.1128/JCM.06664-11
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
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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score 13.070432

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