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

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dc.contributor.authorKamikawa, Camila Mika
dc.contributor.authorMendes, Rinaldo Poncio [UNESP]
dc.contributor.authorVicentini, Adriana Pardini
dc.contributor.institutionCenter of Immunology
dc.contributor.institutionDisease Control Coordination of the São Paulo State Health Secretariat
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2018-12-11T17:09:58Z
dc.date.available2018-12-11T17:09:58Z
dc.date.issued2017-02-15
dc.description.abstractBackground: 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.en
dc.description.affiliationAdolfo Lutz Institute Laboratory of Mycosis Immunodiagnosis Center of Immunology, Av. Dr. Arnaldo, 355, 11o andar
dc.description.affiliationGraduate Program in Sciences Disease Control Coordination of the São Paulo State Health Secretariat
dc.description.affiliationSão Paulo State University (UNESP - Univ Estadual Paulista) Department of Tropical Diseases Botucatu Medical School
dc.description.affiliationUnespSão Paulo State University (UNESP - Univ Estadual Paulista) Department of Tropical Diseases Botucatu Medical School
dc.identifierhttp://dx.doi.org/10.1186/s40409-017-0101-3
dc.identifier.citationJournal of Venomous Animals and Toxins Including Tropical Diseases, v. 23, n. 1, 2017.
dc.identifier.doi10.1186/s40409-017-0101-3
dc.identifier.fileS1678-91992017000100305.pdf
dc.identifier.issn1678-9199
dc.identifier.issn1678-9180
dc.identifier.scieloS1678-91992017000100305
dc.identifier.scopus2-s2.0-85013156386
dc.identifier.urihttp://hdl.handle.net/11449/174240
dc.language.isoeng
dc.relation.ispartofJournal of Venomous Animals and Toxins Including Tropical Diseases
dc.relation.ispartofsjr0,573
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectDot-ELISA
dc.subjectImmunodiagnostic tool
dc.subjectParacoccidioides brasiliensis
dc.subjectParacoccidioidomycosis
dc.subjectValidation
dc.titleStandardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detectionen
dc.typeArtigo

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