The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease
ABSTRACT: Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function...
- Autores:
-
Agudelo Flórez, Piedad
López Quintero, Juan Alvaro
Redher, Jussara
Carneiro Sampaio, Magda
Costa Carvalho, Beatriz
Grumach, Annete
Condino Neto, Antonio
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2004
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/25728
- Acceso en línea:
- http://hdl.handle.net/10495/25728
- Palabra clave:
- Superóxidos
Superoxides
Fagocitos
Phagocytes
Humanos
Human
Neutrófilos
Neutrophils
Estallido Respiratorio
Respiratory Burst
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by/2.5/co/
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UDEA2 |
network_name_str |
Repositorio UdeA |
repository_id_str |
|
dc.title.spa.fl_str_mv |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
title |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
spellingShingle |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease Superóxidos Superoxides Fagocitos Phagocytes Humanos Human Neutrófilos Neutrophils Estallido Respiratorio Respiratory Burst |
title_short |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
title_full |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
title_fullStr |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
title_full_unstemmed |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
title_sort |
The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous disease |
dc.creator.fl_str_mv |
Agudelo Flórez, Piedad López Quintero, Juan Alvaro Redher, Jussara Carneiro Sampaio, Magda Costa Carvalho, Beatriz Grumach, Annete Condino Neto, Antonio |
dc.contributor.author.none.fl_str_mv |
Agudelo Flórez, Piedad López Quintero, Juan Alvaro Redher, Jussara Carneiro Sampaio, Magda Costa Carvalho, Beatriz Grumach, Annete Condino Neto, Antonio |
dc.subject.decs.none.fl_str_mv |
Superóxidos Superoxides Fagocitos Phagocytes Humanos Human Neutrófilos Neutrophils Estallido Respiratorio Respiratory Burst |
topic |
Superóxidos Superoxides Fagocitos Phagocytes Humanos Human Neutrófilos Neutrophils Estallido Respiratorio Respiratory Burst |
description |
ABSTRACT: Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis – a simple, economical and rapid method – was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients. |
publishDate |
2004 |
dc.date.issued.none.fl_str_mv |
2004 |
dc.date.accessioned.none.fl_str_mv |
2022-02-01T16:06:37Z |
dc.date.available.none.fl_str_mv |
2022-02-01T16:06:37Z |
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.issn.none.fl_str_mv |
0100-879X |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/10495/25728 |
dc.identifier.doi.none.fl_str_mv |
10.1590/S0100-879X2004000500001 |
dc.identifier.eissn.none.fl_str_mv |
1414-431X |
identifier_str_mv |
0100-879X 10.1590/S0100-879X2004000500001 1414-431X |
url |
http://hdl.handle.net/10495/25728 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Braz. J. Med. Biol. 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 |
Associação Brasileira de Divulgação Científica |
dc.publisher.group.spa.fl_str_mv |
Inmunodeficiencias Primarias |
dc.publisher.place.spa.fl_str_mv |
São Paulo, Brasil |
institution |
Universidad de Antioquia |
bitstream.url.fl_str_mv |
http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/2/license_rdf http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/1/AgudeloPiedad_2004_UseReverseTranscriptionPCR.pdf http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/3/license.txt |
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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_ |
1812173220881629184 |
spelling |
Agudelo Flórez, PiedadLópez Quintero, Juan AlvaroRedher, JussaraCarneiro Sampaio, MagdaCosta Carvalho, BeatrizGrumach, AnneteCondino Neto, Antonio2022-02-01T16:06:37Z2022-02-01T16:06:37Z20040100-879Xhttp://hdl.handle.net/10495/2572810.1590/S0100-879X20040005000011414-431XABSTRACT: Chronic granulomatous disease (CGD) is an inherited disorder of the innate immune system characterized by a defective oxidative burst of phagocytes and subsequent impairment of their microbicidal activity. Mutations in one of the NADPH-oxidase components affect gene expression or function of this system, leading to the phenotype of CGD. Defects in gp91-phox lead to X-linked CGD, responsible for approximately 70% of CGD cases. Investigation of the highly heterogeneous genotype of CGD patients includes mutation analysis, Northern blot or Western blot assays according to the particular case. The aim of the present study was to use reverse transcription (RT)-PCR for the analysis of molecular defects responsible for X-linked CGD in eight Brazilian patients and to assess its potential for broader application to molecular screening in CGD. Total RNA was prepared from Epstein B virus-transformed B-lymphocytes and reverse transcribed using random hexamers. The resulting cDNA was PCR-amplified by specific and overlapping pairs of primers designed to amplify three regions of the gp91-phox gene: exons 1-5, 3-9, and 7-13. This strategy detected defective gp91-phox expression in seven patients. The RT-PCR results matched clinical history, biochemical data (nitroblue tetrazolium or superoxide release assay) and available mutation analysis in four cases. In three additional cases, RT-PCR results matched clinical history and biochemical data. In another case, RT-PCR was normal despite a clinical history compatible with CGD and defective respiratory burst. We conclude that this new application of RT-PCR analysis – a simple, economical and rapid method – was appropriate for screening molecular defects in 7 of 8 X-linked CGD patients.COL001242610application/pdfengAssociação Brasileira de Divulgação CientíficaInmunodeficiencias PrimariasSão Paulo, Brasilinfo: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/The use of reverse transcription-PCR for the diagnosis of X-linked chronic granulomatous diseaseSuperóxidosSuperoxidesFagocitosPhagocytesHumanosHumanNeutrófilosNeutrophilsEstallido RespiratorioRespiratory BurstBraz. J. Med. Biol. Res.Brazilian Journal of Medical and Biological Research625634375CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8927http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/2/license_rdf1646d1f6b96dbbbc38035efc9239ac9cMD52ORIGINALAgudeloPiedad_2004_UseReverseTranscriptionPCR.pdfAgudeloPiedad_2004_UseReverseTranscriptionPCR.pdfArticulo de investigaciónapplication/pdf846443http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/1/AgudeloPiedad_2004_UseReverseTranscriptionPCR.pdfecaf49ffe1120737e3aae43c427dc2d8MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/25728/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/25728oai:bibliotecadigital.udea.edu.co:10495/257282022-02-01 11:06:38.15Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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 |