Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria
ABSTRACT : Aim: Slow growth rate in culture renders the traditional isolation, identification, and drug susceptibility testing of clinically important mycobacteria inadequate when there is an urgent need for a precise diagnosis in order to initiate patient treatment. Molecular methods all rely on my...
- Autores:
-
Gómez Tangarife, Verónica
Guzmán, Ángela
Mejía, Gloria
Cáceres Contreras, Diego Hernando
Robledo Restrepo, Jaime
Rouzaud, Francois
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2015
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/21043
- Acceso en línea:
- http://hdl.handle.net/10495/21043
- Palabra clave:
- Mycobacteriaceae
Mycobacterium
PCR
Diagnóstico
Diagnosis
Extraccion ADN
PCR (Reacción en cadena de la polimerasa) - Aplicaciones
http://aims.fao.org/aos/agrovoc/c_5018
http://aims.fao.org/aos/agrovoc/c_34079
http://aims.fao.org/aos/agrovoc/c_2238
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by/2.5/co/
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oai:bibliotecadigital.udea.edu.co:10495/21043 |
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Repositorio UdeA |
repository_id_str |
|
dc.title.spa.fl_str_mv |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
title |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
spellingShingle |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria Mycobacteriaceae Mycobacterium PCR Diagnóstico Diagnosis Extraccion ADN PCR (Reacción en cadena de la polimerasa) - Aplicaciones http://aims.fao.org/aos/agrovoc/c_5018 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_2238 |
title_short |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
title_full |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
title_fullStr |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
title_full_unstemmed |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
title_sort |
Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteria |
dc.creator.fl_str_mv |
Gómez Tangarife, Verónica Guzmán, Ángela Mejía, Gloria Cáceres Contreras, Diego Hernando Robledo Restrepo, Jaime Rouzaud, Francois |
dc.contributor.author.none.fl_str_mv |
Gómez Tangarife, Verónica Guzmán, Ángela Mejía, Gloria Cáceres Contreras, Diego Hernando Robledo Restrepo, Jaime Rouzaud, Francois |
dc.subject.decs.none.fl_str_mv |
Mycobacteriaceae |
topic |
Mycobacteriaceae Mycobacterium PCR Diagnóstico Diagnosis Extraccion ADN PCR (Reacción en cadena de la polimerasa) - Aplicaciones http://aims.fao.org/aos/agrovoc/c_5018 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_2238 |
dc.subject.agrovoc.none.fl_str_mv |
Mycobacterium PCR Diagnóstico Diagnosis |
dc.subject.proposal.spa.fl_str_mv |
Extraccion ADN PCR (Reacción en cadena de la polimerasa) - Aplicaciones |
dc.subject.agrovocuri.none.fl_str_mv |
http://aims.fao.org/aos/agrovoc/c_5018 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_2238 |
description |
ABSTRACT : Aim: Slow growth rate in culture renders the traditional isolation, identification, and drug susceptibility testing of clinically important mycobacteria inadequate when there is an urgent need for a precise diagnosis in order to initiate patient treatment. Molecular methods all rely on mycobacterial DNA isolation which in turn has become an essential step of the process. Our study aimed to evaluate DNA isolation protocols from mycobacteria of clinical interest. Methods: Therefore, in order to determine an optimal method we evaluated 8 inexpensive, rapid and easy DNA isolation methods from 30 mycobacterial cultures (10 Mycobacterium tuberculosis and 20 Non-tuberculous Mycobacteria) for subsequent direct detection by PCR. Results: Six of those 8 methods reliably allow the isolation of good DNA yields and quality, the optimal protocol being the one that includes a 1% Triton X-100 lysis solution. Protocols using SDS 1% as a lysis solution did not yield DNA suitable for PCR amplification. Conclusion: Six of the methods we evaluated can easily be implemented in resource limited settings for routine use, potentially contributing to a better management of mycobacterial infections. |
publishDate |
2015 |
dc.date.issued.none.fl_str_mv |
2015 |
dc.date.accessioned.none.fl_str_mv |
2021-07-22T04:40:09Z |
dc.date.available.none.fl_str_mv |
2021-07-22T04:40:09Z |
dc.type.spa.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.hasversion.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/ART |
dc.type.local.spa.fl_str_mv |
Artículo de investigación |
format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
status_str |
publishedVersion |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/10495/21043 |
dc.identifier.doi.none.fl_str_mv |
10.9734/BJMMR/2015/16945 |
dc.identifier.eissn.none.fl_str_mv |
2231-0614 |
url |
http://hdl.handle.net/10495/21043 |
identifier_str_mv |
10.9734/BJMMR/2015/16945 2231-0614 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Br J Med Med Res |
dc.rights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by/2.5/co/ |
dc.rights.accessrights.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
dc.rights.creativecommons.spa.fl_str_mv |
https://creativecommons.org/licenses/by/4.0/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by/2.5/co/ http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by/4.0/ |
dc.format.extent.spa.fl_str_mv |
10 |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.publisher.spa.fl_str_mv |
Sciencedomain International |
dc.publisher.group.spa.fl_str_mv |
Micología Médica y Experimental |
dc.publisher.place.spa.fl_str_mv |
Gurgaon, India |
institution |
Universidad de Antioquia |
bitstream.url.fl_str_mv |
http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/2/license_rdf http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/1/GomezVeronica_2015_Evaluation_Simple_DNA.pdf http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/3/license.txt |
bitstream.checksum.fl_str_mv |
1646d1f6b96dbbbc38035efc9239ac9c e0fa190a2a1c50c879a98eb29853021f 8a4605be74aa9ea9d79846c1fba20a33 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositorio Institucional Universidad de Antioquia |
repository.mail.fl_str_mv |
andres.perez@udea.edu.co |
_version_ |
1812173163971215360 |
spelling |
Gómez Tangarife, VerónicaGuzmán, ÁngelaMejía, GloriaCáceres Contreras, Diego HernandoRobledo Restrepo, JaimeRouzaud, Francois2021-07-22T04:40:09Z2021-07-22T04:40:09Z2015http://hdl.handle.net/10495/2104310.9734/BJMMR/2015/169452231-0614ABSTRACT : Aim: Slow growth rate in culture renders the traditional isolation, identification, and drug susceptibility testing of clinically important mycobacteria inadequate when there is an urgent need for a precise diagnosis in order to initiate patient treatment. Molecular methods all rely on mycobacterial DNA isolation which in turn has become an essential step of the process. Our study aimed to evaluate DNA isolation protocols from mycobacteria of clinical interest. Methods: Therefore, in order to determine an optimal method we evaluated 8 inexpensive, rapid and easy DNA isolation methods from 30 mycobacterial cultures (10 Mycobacterium tuberculosis and 20 Non-tuberculous Mycobacteria) for subsequent direct detection by PCR. Results: Six of those 8 methods reliably allow the isolation of good DNA yields and quality, the optimal protocol being the one that includes a 1% Triton X-100 lysis solution. Protocols using SDS 1% as a lysis solution did not yield DNA suitable for PCR amplification. Conclusion: Six of the methods we evaluated can easily be implemented in resource limited settings for routine use, potentially contributing to a better management of mycobacterial infections.COL001370910application/pdfengSciencedomain InternationalMicología Médica y ExperimentalGurgaon, Indiainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by/4.0/Evaluation of simple and Cost-Effective DNA preparation and subsequent PCR amplification for clinically relevant mycobacteriaMycobacteriaceaeMycobacteriumPCRDiagnósticoDiagnosisExtraccion ADNPCR (Reacción en cadena de la polimerasa) - Aplicacioneshttp://aims.fao.org/aos/agrovoc/c_5018http://aims.fao.org/aos/agrovoc/c_34079http://aims.fao.org/aos/agrovoc/c_2238Br J Med Med ResBritish Journal Of Medicine And Medical Research14715682CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8927http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/2/license_rdf1646d1f6b96dbbbc38035efc9239ac9cMD52ORIGINALGomezVeronica_2015_Evaluation_Simple_DNA.pdfGomezVeronica_2015_Evaluation_Simple_DNA.pdfArtículo de investigaciónapplication/pdf427806http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/1/GomezVeronica_2015_Evaluation_Simple_DNA.pdfe0fa190a2a1c50c879a98eb29853021fMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/21043/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/21043oai:bibliotecadigital.udea.edu.co:10495/210432022-04-22 10:18:50.86Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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 |