A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Outros Autores: | , , , , , , |
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/166329 |
Resumo: | Candidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans, spiked human blood, the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/µL), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/µL). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R2 ) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting. |
id |
IMT-1_02707edfed64a6f6fd654b0d8d2104b0 |
---|---|
oai_identifier_str |
oai:revistas.usp.br:article/166329 |
network_acronym_str |
IMT-1 |
network_name_str |
Revista do Instituto de Medicina Tropical de São Paulo |
repository_id_str |
|
spelling |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human bloodCandida albicansCandidemiaDetectionInvasive candidiasisITS2Quantitative PCRCandidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans, spiked human blood, the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/µL), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/µL). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R2 ) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting.Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo2020-02-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/xmlhttps://www.revistas.usp.br/rimtsp/article/view/16632910.1590/S1678-9946202062009Revista do Instituto de Medicina Tropical de São Paulo; Vol. 62 (2020); e9Revista do Instituto de Medicina Tropical de São Paulo; Vol. 62 (2020); e9Revista do Instituto de Medicina Tropical de São Paulo; v. 62 (2020); e91678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTenghttps://www.revistas.usp.br/rimtsp/article/view/166329/159061https://www.revistas.usp.br/rimtsp/article/view/166329/159062Copyright (c) 2020 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessBusser, Felipe DelatorreCoelho, Vivian CasoFonseca, Claudia de AbreuDel Negro, Gilda Maria BarbaroShikanai-Yasuda, Maria AparecidaLopes, Marta HeloisaMagri, Marcello Mihailenko ChavesFreitas, Vera Lucia Teixeira de2020-10-26T17:39:25Zoai:revistas.usp.br:article/166329Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:52:51.592548Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true |
dc.title.none.fl_str_mv |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
title |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
spellingShingle |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood Busser, Felipe Delatorre Candida albicans Candidemia Detection Invasive candidiasis ITS2 Quantitative PCR |
title_short |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
title_full |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
title_fullStr |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
title_full_unstemmed |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
title_sort |
A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood |
author |
Busser, Felipe Delatorre |
author_facet |
Busser, Felipe Delatorre Coelho, Vivian Caso Fonseca, Claudia de Abreu Del Negro, Gilda Maria Barbaro Shikanai-Yasuda, Maria Aparecida Lopes, Marta Heloisa Magri, Marcello Mihailenko Chaves Freitas, Vera Lucia Teixeira de |
author_role |
author |
author2 |
Coelho, Vivian Caso Fonseca, Claudia de Abreu Del Negro, Gilda Maria Barbaro Shikanai-Yasuda, Maria Aparecida Lopes, Marta Heloisa Magri, Marcello Mihailenko Chaves Freitas, Vera Lucia Teixeira de |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Busser, Felipe Delatorre Coelho, Vivian Caso Fonseca, Claudia de Abreu Del Negro, Gilda Maria Barbaro Shikanai-Yasuda, Maria Aparecida Lopes, Marta Heloisa Magri, Marcello Mihailenko Chaves Freitas, Vera Lucia Teixeira de |
dc.subject.por.fl_str_mv |
Candida albicans Candidemia Detection Invasive candidiasis ITS2 Quantitative PCR |
topic |
Candida albicans Candidemia Detection Invasive candidiasis ITS2 Quantitative PCR |
description |
Candidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans, spiked human blood, the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/µL), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/µL). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R2 ) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-02-04 |
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/166329 10.1590/S1678-9946202062009 |
url |
https://www.revistas.usp.br/rimtsp/article/view/166329 |
identifier_str_mv |
10.1590/S1678-9946202062009 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/166329/159061 https://www.revistas.usp.br/rimtsp/article/view/166329/159062 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2020 Revista do Instituto de Medicina Tropical de São Paulo info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2020 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. 62 (2020); e9 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 62 (2020); e9 Revista do Instituto de Medicina Tropical de São Paulo; v. 62 (2020); e9 1678-9946 0036-4665 reponame:Revista do Instituto de Medicina Tropical de São Paulo instname:Instituto de Medicina Tropical (IMT) instacron:IMT |
instname_str |
Instituto de Medicina Tropical (IMT) |
instacron_str |
IMT |
institution |
IMT |
reponame_str |
Revista do Instituto de Medicina Tropical de São Paulo |
collection |
Revista do Instituto de Medicina Tropical de São Paulo |
repository.name.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT) |
repository.mail.fl_str_mv |
||revimtsp@usp.br |
_version_ |
1798951652662706176 |