Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates

Detalhes bibliográficos
Autor(a) principal: Reyes-Montes, María del Rocío
Data de Publicação: 2019
Outros Autores: Acosta-Altamirano, Gustavo, Duarte-Escalante, Esperanza, Salazar, Eduardo García, Martínez-Herrera, Erick, Arenas, Roberto, González, Gloria, Frías-De-León, María Guadalupe
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Revista do Instituto de Medicina Tropical de São Paulo
Texto Completo: https://www.revistas.usp.br/rimtsp/article/view/162383
Resumo: Candida glabrata complex includes three species identified through molecular biology methods: C. glabrata sensu stricto, C. nivariensis and C. bracarensis. In Mexico, the phenotypic methods are still used in the diagnosis; therefore, the presence of C. nivariensis and C. bracarensis among clinical isolates is still unknown. The aim of this study was to evaluate the utility of a multiplex PCR for the identification of the C. glabrata species complex. DNA samples from 92 clinical isolates that were previously identified through phenotypic characteristics as C. glabrata were amplified by four oligonucleotides (UNI-5.8S, GLA-f, BRA-f, and NIV-f) that generate amplicons of 397, 293 and 223-bp corresponding to C. glabrata sensu stricto, C. nivariensis, and C. bracarensis, respectively. The amplicon sequences were used to perform a phylogenetic analysis through the Maximum Likelihood method (MEGA6), including strains and reference sequences of species belonging to C. glabrata complex. In addition, recombination and linkage disequilibrium were estimated (DnaSP version 5.0) for C. glabrata sensu stricto isolates. Eighty-eight isolates generated a 397-bp fragment and only in one isolate a 223-bp amplicon was observed. In the phylogenetic tree, the sequences of 397-bp were grouped with C. glabrata reference sequences, and the sequence of 223-bp was grouped with C. bracarensis reference sequences, corroborating the PCR identification. The number of recombination events for the isolates of C. glabrata sensu stricto was zero, suggesting a clonal population structure. Three isolates that did not amplify any of the expected fragments were identified as Saccharomyces cerevisiae through the sequencing of the D1/D2 domain region within the 28S rDNA gene. The multiplex PCR is a fast, cost-effective and reliable tool that can be used in clinical laboratories to identify C. glabrata complex species.
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spelling Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolatesCandida glabrata complexCandida bracarensisCandida nivariensisMultiplex PCRCandidiasisMexicoCandida glabrata complex includes three species identified through molecular biology methods: C. glabrata sensu stricto, C. nivariensis and C. bracarensis. In Mexico, the phenotypic methods are still used in the diagnosis; therefore, the presence of C. nivariensis and C. bracarensis among clinical isolates is still unknown. The aim of this study was to evaluate the utility of a multiplex PCR for the identification of the C. glabrata species complex. DNA samples from 92 clinical isolates that were previously identified through phenotypic characteristics as C. glabrata were amplified by four oligonucleotides (UNI-5.8S, GLA-f, BRA-f, and NIV-f) that generate amplicons of 397, 293 and 223-bp corresponding to C. glabrata sensu stricto, C. nivariensis, and C. bracarensis, respectively. The amplicon sequences were used to perform a phylogenetic analysis through the Maximum Likelihood method (MEGA6), including strains and reference sequences of species belonging to C. glabrata complex. In addition, recombination and linkage disequilibrium were estimated (DnaSP version 5.0) for C. glabrata sensu stricto isolates. Eighty-eight isolates generated a 397-bp fragment and only in one isolate a 223-bp amplicon was observed. In the phylogenetic tree, the sequences of 397-bp were grouped with C. glabrata reference sequences, and the sequence of 223-bp was grouped with C. bracarensis reference sequences, corroborating the PCR identification. The number of recombination events for the isolates of C. glabrata sensu stricto was zero, suggesting a clonal population structure. Three isolates that did not amplify any of the expected fragments were identified as Saccharomyces cerevisiae through the sequencing of the D1/D2 domain region within the 28S rDNA gene. The multiplex PCR is a fast, cost-effective and reliable tool that can be used in clinical laboratories to identify C. glabrata complex species.Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo2019-09-18info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/xmlhttps://www.revistas.usp.br/rimtsp/article/view/16238310.1590/s1678-9946201961037Revista do Instituto de Medicina Tropical de São Paulo; Vol. 61 (2019); e37Revista do Instituto de Medicina Tropical de São Paulo; Vol. 61 (2019); e37Revista do Instituto de Medicina Tropical de São Paulo; v. 61 (2019); e371678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTenghttps://www.revistas.usp.br/rimtsp/article/view/162383/156177https://www.revistas.usp.br/rimtsp/article/view/162383/156178Copyright (c) 2019 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessReyes-Montes, María del RocíoAcosta-Altamirano, GustavoDuarte-Escalante, EsperanzaSalazar, Eduardo GarcíaMartínez-Herrera, ErickArenas, RobertoGonzález, GloriaFrías-De-León, María Guadalupe2019-09-18T13:19:42Zoai:revistas.usp.br:article/162383Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:52:49.890631Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true
dc.title.none.fl_str_mv Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
title Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
spellingShingle Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
Reyes-Montes, María del Rocío
Candida glabrata complex
Candida bracarensis
Candida nivariensis
Multiplex PCR
Candidiasis
Mexico
title_short Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
title_full Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
title_fullStr Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
title_full_unstemmed Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
title_sort Usefulness of a multiplex PCR for the rapid identification of Candida glabrata species complex in Mexican clinical isolates
author Reyes-Montes, María del Rocío
author_facet Reyes-Montes, María del Rocío
Acosta-Altamirano, Gustavo
Duarte-Escalante, Esperanza
Salazar, Eduardo García
Martínez-Herrera, Erick
Arenas, Roberto
González, Gloria
Frías-De-León, María Guadalupe
author_role author
author2 Acosta-Altamirano, Gustavo
Duarte-Escalante, Esperanza
Salazar, Eduardo García
Martínez-Herrera, Erick
Arenas, Roberto
González, Gloria
Frías-De-León, María Guadalupe
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Reyes-Montes, María del Rocío
Acosta-Altamirano, Gustavo
Duarte-Escalante, Esperanza
Salazar, Eduardo García
Martínez-Herrera, Erick
Arenas, Roberto
González, Gloria
Frías-De-León, María Guadalupe
dc.subject.por.fl_str_mv Candida glabrata complex
Candida bracarensis
Candida nivariensis
Multiplex PCR
Candidiasis
Mexico
topic Candida glabrata complex
Candida bracarensis
Candida nivariensis
Multiplex PCR
Candidiasis
Mexico
description Candida glabrata complex includes three species identified through molecular biology methods: C. glabrata sensu stricto, C. nivariensis and C. bracarensis. In Mexico, the phenotypic methods are still used in the diagnosis; therefore, the presence of C. nivariensis and C. bracarensis among clinical isolates is still unknown. The aim of this study was to evaluate the utility of a multiplex PCR for the identification of the C. glabrata species complex. DNA samples from 92 clinical isolates that were previously identified through phenotypic characteristics as C. glabrata were amplified by four oligonucleotides (UNI-5.8S, GLA-f, BRA-f, and NIV-f) that generate amplicons of 397, 293 and 223-bp corresponding to C. glabrata sensu stricto, C. nivariensis, and C. bracarensis, respectively. The amplicon sequences were used to perform a phylogenetic analysis through the Maximum Likelihood method (MEGA6), including strains and reference sequences of species belonging to C. glabrata complex. In addition, recombination and linkage disequilibrium were estimated (DnaSP version 5.0) for C. glabrata sensu stricto isolates. Eighty-eight isolates generated a 397-bp fragment and only in one isolate a 223-bp amplicon was observed. In the phylogenetic tree, the sequences of 397-bp were grouped with C. glabrata reference sequences, and the sequence of 223-bp was grouped with C. bracarensis reference sequences, corroborating the PCR identification. The number of recombination events for the isolates of C. glabrata sensu stricto was zero, suggesting a clonal population structure. Three isolates that did not amplify any of the expected fragments were identified as Saccharomyces cerevisiae through the sequencing of the D1/D2 domain region within the 28S rDNA gene. The multiplex PCR is a fast, cost-effective and reliable tool that can be used in clinical laboratories to identify C. glabrata complex species.
publishDate 2019
dc.date.none.fl_str_mv 2019-09-18
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://www.revistas.usp.br/rimtsp/article/view/162383
10.1590/s1678-9946201961037
url https://www.revistas.usp.br/rimtsp/article/view/162383
identifier_str_mv 10.1590/s1678-9946201961037
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://www.revistas.usp.br/rimtsp/article/view/162383/156177
https://www.revistas.usp.br/rimtsp/article/view/162383/156178
dc.rights.driver.fl_str_mv Copyright (c) 2019 Revista do Instituto de Medicina Tropical de São Paulo
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Copyright (c) 2019 Revista do Instituto de Medicina Tropical de São Paulo
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/xml
dc.publisher.none.fl_str_mv Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo
publisher.none.fl_str_mv Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo
dc.source.none.fl_str_mv Revista do Instituto de Medicina Tropical de São Paulo; Vol. 61 (2019); e37
Revista do Instituto de Medicina Tropical de São Paulo; Vol. 61 (2019); e37
Revista do Instituto de Medicina Tropical de São Paulo; v. 61 (2019); e37
1678-9946
0036-4665
reponame:Revista do Instituto de Medicina Tropical de São Paulo
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