Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , |
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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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) |
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1808128196181229568 |