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...

Full description

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/
id UDEA2_65374438ede6fda0b356f4724804a7f3
oai_identifier_str oai:bibliotecadigital.udea.edu.co:10495/31149
network_acronym_str UDEA2
network_name_str 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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