Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection

Detalhes bibliográficos
Autor(a) principal: Kamikawa, Camila Mika
Data de Publicação: 2017
Outros Autores: Mendes, Rinaldo Poncio [UNESP], Vicentini, Adriana Pardini
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1186/s40409-017-0101-3
http://hdl.handle.net/11449/174240
Resumo: Background: Paracoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI). Methods: In order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI. Results: The DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization. Conclusion: Dot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life - membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories - and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.
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spelling Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detectionDot-ELISAImmunodiagnostic toolParacoccidioides brasiliensisParacoccidioidomycosisValidationBackground: Paracoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI). Methods: In order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI. Results: The DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization. Conclusion: Dot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life - membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories - and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.Adolfo Lutz Institute Laboratory of Mycosis Immunodiagnosis Center of Immunology, Av. Dr. Arnaldo, 355, 11o andarGraduate Program in Sciences Disease Control Coordination of the São Paulo State Health SecretariatSão Paulo State University (UNESP - Univ Estadual Paulista) Department of Tropical Diseases Botucatu Medical SchoolSão Paulo State University (UNESP - Univ Estadual Paulista) Department of Tropical Diseases Botucatu Medical SchoolCenter of ImmunologyDisease Control Coordination of the São Paulo State Health SecretariatUniversidade Estadual Paulista (Unesp)Kamikawa, Camila MikaMendes, Rinaldo Poncio [UNESP]Vicentini, Adriana Pardini2018-12-11T17:09:58Z2018-12-11T17:09:58Z2017-02-15info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1186/s40409-017-0101-3Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.1678-91991678-9180http://hdl.handle.net/11449/17424010.1186/s40409-017-0101-3S1678-919920170001003052-s2.0-85013156386S1678-91992017000100305.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Venomous Animals and Toxins Including Tropical Diseases0,573info:eu-repo/semantics/openAccess2024-08-15T15:23:16Zoai:repositorio.unesp.br:11449/174240Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-15T15:23:16Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
title Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
spellingShingle Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
Kamikawa, Camila Mika
Dot-ELISA
Immunodiagnostic tool
Paracoccidioides brasiliensis
Paracoccidioidomycosis
Validation
title_short Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
title_full Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
title_fullStr Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
title_full_unstemmed Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
title_sort Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
author Kamikawa, Camila Mika
author_facet Kamikawa, Camila Mika
Mendes, Rinaldo Poncio [UNESP]
Vicentini, Adriana Pardini
author_role author
author2 Mendes, Rinaldo Poncio [UNESP]
Vicentini, Adriana Pardini
author2_role author
author
dc.contributor.none.fl_str_mv Center of Immunology
Disease Control Coordination of the São Paulo State Health Secretariat
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Kamikawa, Camila Mika
Mendes, Rinaldo Poncio [UNESP]
Vicentini, Adriana Pardini
dc.subject.por.fl_str_mv Dot-ELISA
Immunodiagnostic tool
Paracoccidioides brasiliensis
Paracoccidioidomycosis
Validation
topic Dot-ELISA
Immunodiagnostic tool
Paracoccidioides brasiliensis
Paracoccidioidomycosis
Validation
description Background: Paracoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI). Methods: In order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI. Results: The DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization. Conclusion: Dot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life - membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories - and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.
publishDate 2017
dc.date.none.fl_str_mv 2017-02-15
2018-12-11T17:09:58Z
2018-12-11T17:09:58Z
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/10.1186/s40409-017-0101-3
Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.
1678-9199
1678-9180
http://hdl.handle.net/11449/174240
10.1186/s40409-017-0101-3
S1678-91992017000100305
2-s2.0-85013156386
S1678-91992017000100305.pdf
url http://dx.doi.org/10.1186/s40409-017-0101-3
http://hdl.handle.net/11449/174240
identifier_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.
1678-9199
1678-9180
10.1186/s40409-017-0101-3
S1678-91992017000100305
2-s2.0-85013156386
S1678-91992017000100305.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases
0,573
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 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
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