PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies
ABSTRACT: The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis gene...
- Autores:
-
Díez Posada, Soraya
Pino Tamayo, Paula Andrea
Corredor Rengifo, Gabriel German
Castaño, John Harold
Peralta, Luis Alejandro
McEwen Ochoa, Juan Guillermo
Restrepo Moreno, Ángela
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 1999
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/31149
- Acceso en línea:
- https://hdl.handle.net/10495/31149
- Palabra clave:
- Paracoccidioides
Reacción en Cadena de la Polimerasa
Polymerase Chain Reaction
Cartilla de ADN
DNA Primers
Northern Blotting
Blotting, Northern
ADN de Hongos
DNA, Fungal
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by-nc-sa/2.5/co/
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UDEA2 |
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Repositorio UdeA |
repository_id_str |
|
dc.title.spa.fl_str_mv |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
title |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
spellingShingle |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies Paracoccidioides Reacción en Cadena de la Polimerasa Polymerase Chain Reaction Cartilla de ADN DNA Primers Northern Blotting Blotting, Northern ADN de Hongos DNA, Fungal |
title_short |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
title_full |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
title_fullStr |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
title_full_unstemmed |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
title_sort |
PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological Studies |
dc.creator.fl_str_mv |
Díez Posada, Soraya Pino Tamayo, Paula Andrea Corredor Rengifo, Gabriel German Castaño, John Harold Peralta, Luis Alejandro McEwen Ochoa, Juan Guillermo Restrepo Moreno, Ángela |
dc.contributor.author.none.fl_str_mv |
Díez Posada, Soraya Pino Tamayo, Paula Andrea Corredor Rengifo, Gabriel German Castaño, John Harold Peralta, Luis Alejandro McEwen Ochoa, Juan Guillermo Restrepo Moreno, Ángela |
dc.subject.decs.none.fl_str_mv |
Paracoccidioides Reacción en Cadena de la Polimerasa Polymerase Chain Reaction Cartilla de ADN DNA Primers Northern Blotting Blotting, Northern ADN de Hongos DNA, Fungal |
topic |
Paracoccidioides Reacción en Cadena de la Polimerasa Polymerase Chain Reaction Cartilla de ADN DNA Primers Northern Blotting Blotting, Northern ADN de Hongos DNA, Fungal |
description |
ABSTRACT: The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis genes. This fungus as well as several other fungi, were grown and their DNA obtained by mechanical disruption and a phenol chloroform isoamylalcohol-based purification method. The DNA served for a PCR reaction that employed specific primers from two P. brasiliensis genes that codify for antigenic proteins, namely, the 27 kDa and the 43 kDa. The lowest detection range for the 27 kDa gene was 3 pg. The amplification for both genes was positive only with DNA from P. brasiliensis; additionally, the mRNA for the 27 kDa gene was present only in P. brasiliensis, as indicated by the Northern analysis. The standardization of PCR technology permitted the amplification of P. brasiliensis DNA in artificially contaminated soils and in tissues of armadillos naturally infected with the fungus. These results indicate that PCR technology could play an important role in the search for P. brasiliensis’ habitat and could also be used in other ecological studies. |
publishDate |
1999 |
dc.date.issued.none.fl_str_mv |
1999 |
dc.date.accessioned.none.fl_str_mv |
2022-10-08T18:19:01Z |
dc.date.available.none.fl_str_mv |
2022-10-08T18:19:01Z |
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.citation.spa.fl_str_mv |
Díez S, Garcia EA, Pino PA, Botero S, Corredor GG, Peralta LA, Castaño JH, Restrepo A, McEwen JG. PCR with Paracoccidioides brasiliensis specific primers: potential use in ecological studies. Rev Inst Med Trop Sao Paulo. 1999 Nov-Dec;41(6):351-8. doi: 10.1590/s0036-46651999000600004. |
dc.identifier.issn.none.fl_str_mv |
0036-4665 |
dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/10495/31149 |
dc.identifier.doi.none.fl_str_mv |
10.1590/S0036-46651999000600004 |
dc.identifier.eissn.none.fl_str_mv |
1678-9946 |
identifier_str_mv |
Díez S, Garcia EA, Pino PA, Botero S, Corredor GG, Peralta LA, Castaño JH, Restrepo A, McEwen JG. PCR with Paracoccidioides brasiliensis specific primers: potential use in ecological studies. Rev Inst Med Trop Sao Paulo. 1999 Nov-Dec;41(6):351-8. doi: 10.1590/s0036-46651999000600004. 0036-4665 10.1590/S0036-46651999000600004 1678-9946 |
url |
https://hdl.handle.net/10495/31149 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Rev. Inst. Med. Trop. São Paulo. |
dc.rights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by-nc-sa/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-nc-sa/4.0/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-sa/2.5/co/ http://purl.org/coar/access_right/c_abf2 https://creativecommons.org/licenses/by-nc-sa/4.0/ |
dc.format.extent.spa.fl_str_mv |
7 |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.publisher.spa.fl_str_mv |
Universidade de São Paulo, Faculdade de Medicina |
dc.publisher.group.spa.fl_str_mv |
Micología Médica y Experimental |
dc.publisher.place.spa.fl_str_mv |
Sao Paulo, Brasil |
institution |
Universidad de Antioquia |
bitstream.url.fl_str_mv |
https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/1/RestrepoAngela_1999_PCR_Paracoccidioides_brasiliensis.pdf https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/2/license_rdf https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/3/license.txt |
bitstream.checksum.fl_str_mv |
35529b8dfb19ea8e0f92e8f13feab824 e2060682c9c70d4d30c83c51448f4eed 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_ |
1812173130458726400 |
spelling |
Díez Posada, SorayaPino Tamayo, Paula AndreaCorredor Rengifo, Gabriel GermanCastaño, John HaroldPeralta, Luis AlejandroMcEwen Ochoa, Juan GuillermoRestrepo Moreno, Ángela2022-10-08T18:19:01Z2022-10-08T18:19:01Z1999Díez S, Garcia EA, Pino PA, Botero S, Corredor GG, Peralta LA, Castaño JH, Restrepo A, McEwen JG. PCR with Paracoccidioides brasiliensis specific primers: potential use in ecological studies. Rev Inst Med Trop Sao Paulo. 1999 Nov-Dec;41(6):351-8. doi: 10.1590/s0036-46651999000600004.0036-4665https://hdl.handle.net/10495/3114910.1590/S0036-466519990006000041678-9946ABSTRACT: The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis genes. This fungus as well as several other fungi, were grown and their DNA obtained by mechanical disruption and a phenol chloroform isoamylalcohol-based purification method. The DNA served for a PCR reaction that employed specific primers from two P. brasiliensis genes that codify for antigenic proteins, namely, the 27 kDa and the 43 kDa. The lowest detection range for the 27 kDa gene was 3 pg. The amplification for both genes was positive only with DNA from P. brasiliensis; additionally, the mRNA for the 27 kDa gene was present only in P. brasiliensis, as indicated by the Northern analysis. The standardization of PCR technology permitted the amplification of P. brasiliensis DNA in artificially contaminated soils and in tissues of armadillos naturally infected with the fungus. These results indicate that PCR technology could play an important role in the search for P. brasiliensis’ habitat and could also be used in other ecological studies.COL00137097application/pdfengUniversidade de São Paulo, Faculdade de MedicinaMicología Médica y ExperimentalSao 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-nc-sa/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by-nc-sa/4.0/PCR with Paracoccidioides brasiliensis Specific Primers : Potential Use in Ecological StudiesParacoccidioidesReacción en Cadena de la PolimerasaPolymerase Chain ReactionCartilla de ADNDNA PrimersNorthern BlottingBlotting, NorthernADN de HongosDNA, FungalRev. Inst. Med. Trop. São Paulo.Revista do Instituto de Medicina Tropical de Sao Paulo351357416ORIGINALRestrepoAngela_1999_PCR_Paracoccidioides_brasiliensis.pdfRestrepoAngela_1999_PCR_Paracoccidioides_brasiliensis.pdfArtículo de investigaciónapplication/pdf217887https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/1/RestrepoAngela_1999_PCR_Paracoccidioides_brasiliensis.pdf35529b8dfb19ea8e0f92e8f13feab824MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-81051https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/2/license_rdfe2060682c9c70d4d30c83c51448f4eedMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://bibliotecadigital.udea.edu.co/bitstream/10495/31149/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/31149oai:bibliotecadigital.udea.edu.co:10495/311492022-10-08 13:19:02.616Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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 |