Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1093/mmy/myv103 http://hdl.handle.net/11449/172870 |
Resumo: | The aim of this study was to establish a reproducible protocol using the methodology of hyaline zones around the colonies on specific agar plates for phospholipase and proteinase production. This was an in vitro double-blind experiment, in which the dependent variables were the enzymatic activity measurements (Pz) for the production of phospholipase (Pz-ph) and the production of secreted aspartyl proteinases (Pz-sap). Three independent variables give rise to different measurement protocols. All measurements were carried out at two different moments by four examiners (E1, E2, E3, and E4). The minimum sample size was 30 Candida albicans clinical isolates. Specific agar plates for phospholipase and SAPs production were prepared according the literature. The intra-and inter-examiner reproducibility for each protocol was estimated using the Intraclass Correlation Coefficient (ICC) and its confidence interval (95% CI). Based on the results obtained for both phospholipase and SAPs, there appears to be no consensus on the protocol chosen for each particular examiner. Measuring the colonies in triplicate may be the main factor associated with the increase in measurement accuracy and should therefore take precedence over measuring only one colony. When only one examiner is responsible for taking measurements, a standard protocol should be put in place and the statistical calibration of this researcher should be done prior to data collection. However, if two or more researchers are involved in the assessment of agar plates, our results suggest that the protocols using software to undertake plate reading is preferred. |
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Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicansCandida albicansReproducibility of ResultsVirulence factorsThe aim of this study was to establish a reproducible protocol using the methodology of hyaline zones around the colonies on specific agar plates for phospholipase and proteinase production. This was an in vitro double-blind experiment, in which the dependent variables were the enzymatic activity measurements (Pz) for the production of phospholipase (Pz-ph) and the production of secreted aspartyl proteinases (Pz-sap). Three independent variables give rise to different measurement protocols. All measurements were carried out at two different moments by four examiners (E1, E2, E3, and E4). The minimum sample size was 30 Candida albicans clinical isolates. Specific agar plates for phospholipase and SAPs production were prepared according the literature. The intra-and inter-examiner reproducibility for each protocol was estimated using the Intraclass Correlation Coefficient (ICC) and its confidence interval (95% CI). Based on the results obtained for both phospholipase and SAPs, there appears to be no consensus on the protocol chosen for each particular examiner. Measuring the colonies in triplicate may be the main factor associated with the increase in measurement accuracy and should therefore take precedence over measuring only one colony. When only one examiner is responsible for taking measurements, a standard protocol should be put in place and the statistical calibration of this researcher should be done prior to data collection. However, if two or more researchers are involved in the assessment of agar plates, our results suggest that the protocols using software to undertake plate reading is preferred.Department of Social Dentistry Araraquara Dental School UNESP-Univ Estadual Paulista, Humaitá Street 1680Department of Dental Materials and Prosthodontics Araraquara Dental School UNESP-Univ Estadual Paulista, Humaitá Street 1680Department of Social Dentistry Araraquara Dental School UNESP-Univ Estadual Paulista, Humaitá Street 1680Department of Dental Materials and Prosthodontics Araraquara Dental School UNESP-Univ Estadual Paulista, Humaitá Street 1680Universidade Estadual Paulista (Unesp)Arantes, Paula Tamiaõ [UNESP]Sanitá, Paula Volpato [UNESP]Santezi, Carolina [UNESP]Barbeiro, Camila De Oliveira [UNESP]Reina, Bárbara Donadon [UNESP]Vergani, Carlos Eduardo [UNESP]Dovigo, Lívia Nordi [UNESP]2018-12-11T17:02:30Z2018-12-11T17:02:30Z2016-01-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article266-274application/pdfhttp://dx.doi.org/10.1093/mmy/myv103Medical Mycology, v. 54, n. 3, p. 266-274, 2016.1460-27091369-3786http://hdl.handle.net/11449/17287010.1093/mmy/myv1032-s2.0-849644130192-s2.0-84964413019.pdf0127570480681342Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMedical Mycology0,9730,973info:eu-repo/semantics/openAccess2023-10-12T06:07:46Zoai:repositorio.unesp.br:11449/172870Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-12T06:07:46Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
title |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
spellingShingle |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans Arantes, Paula Tamiaõ [UNESP] Candida albicans Reproducibility of Results Virulence factors |
title_short |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
title_full |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
title_fullStr |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
title_full_unstemmed |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
title_sort |
Reliability of the agar based method to assess the production of degradative enzymes in clinical isolates of Candida albicans |
author |
Arantes, Paula Tamiaõ [UNESP] |
author_facet |
Arantes, Paula Tamiaõ [UNESP] Sanitá, Paula Volpato [UNESP] Santezi, Carolina [UNESP] Barbeiro, Camila De Oliveira [UNESP] Reina, Bárbara Donadon [UNESP] Vergani, Carlos Eduardo [UNESP] Dovigo, Lívia Nordi [UNESP] |
author_role |
author |
author2 |
Sanitá, Paula Volpato [UNESP] Santezi, Carolina [UNESP] Barbeiro, Camila De Oliveira [UNESP] Reina, Bárbara Donadon [UNESP] Vergani, Carlos Eduardo [UNESP] Dovigo, Lívia Nordi [UNESP] |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Arantes, Paula Tamiaõ [UNESP] Sanitá, Paula Volpato [UNESP] Santezi, Carolina [UNESP] Barbeiro, Camila De Oliveira [UNESP] Reina, Bárbara Donadon [UNESP] Vergani, Carlos Eduardo [UNESP] Dovigo, Lívia Nordi [UNESP] |
dc.subject.por.fl_str_mv |
Candida albicans Reproducibility of Results Virulence factors |
topic |
Candida albicans Reproducibility of Results Virulence factors |
description |
The aim of this study was to establish a reproducible protocol using the methodology of hyaline zones around the colonies on specific agar plates for phospholipase and proteinase production. This was an in vitro double-blind experiment, in which the dependent variables were the enzymatic activity measurements (Pz) for the production of phospholipase (Pz-ph) and the production of secreted aspartyl proteinases (Pz-sap). Three independent variables give rise to different measurement protocols. All measurements were carried out at two different moments by four examiners (E1, E2, E3, and E4). The minimum sample size was 30 Candida albicans clinical isolates. Specific agar plates for phospholipase and SAPs production were prepared according the literature. The intra-and inter-examiner reproducibility for each protocol was estimated using the Intraclass Correlation Coefficient (ICC) and its confidence interval (95% CI). Based on the results obtained for both phospholipase and SAPs, there appears to be no consensus on the protocol chosen for each particular examiner. Measuring the colonies in triplicate may be the main factor associated with the increase in measurement accuracy and should therefore take precedence over measuring only one colony. When only one examiner is responsible for taking measurements, a standard protocol should be put in place and the statistical calibration of this researcher should be done prior to data collection. However, if two or more researchers are involved in the assessment of agar plates, our results suggest that the protocols using software to undertake plate reading is preferred. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-01-14 2018-12-11T17:02:30Z 2018-12-11T17:02:30Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1093/mmy/myv103 Medical Mycology, v. 54, n. 3, p. 266-274, 2016. 1460-2709 1369-3786 http://hdl.handle.net/11449/172870 10.1093/mmy/myv103 2-s2.0-84964413019 2-s2.0-84964413019.pdf 0127570480681342 |
url |
http://dx.doi.org/10.1093/mmy/myv103 http://hdl.handle.net/11449/172870 |
identifier_str_mv |
Medical Mycology, v. 54, n. 3, p. 266-274, 2016. 1460-2709 1369-3786 10.1093/mmy/myv103 2-s2.0-84964413019 2-s2.0-84964413019.pdf 0127570480681342 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Medical Mycology 0,973 0,973 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
266-274 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1799964534203482112 |