Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Detalhes bibliográficos
Autor(a) principal: Tomazini, Karina Andressa [UNESP]
Data de Publicação: 2023
Outros Autores: Pereira, Beatriz Aparecida Soares [UNESP], Sylvestre, Tatiane Fernanda, de Souza Cavalcante, Ricardo [UNESP], de Carvalho, Lídia Raquel [UNESP], Mendes, Rinaldo Poncio [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/1678-9199-JVATITD-2022-0045
http://hdl.handle.net/11449/248497
Resumo: Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at –80oC for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at –20oC for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at –80oC for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at –20oC.
id UNSP_a8317dac9b14fec28c62e16e81a15559
oai_identifier_str oai:repositorio.unesp.br:11449/248497
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patientsBlood matricesParacoccidioidomycosisPlasmaSerological diagnosisSerumStorageBackground: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at –80oC for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at –20oC for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at –80oC for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at –20oC.Universidade Estadual PaulistaDepartment of Infectology Dermatology Diagnostic Imaging and Radiotherapy Botucatu Medical School (FMB) São Paulo State University (UNESP), SPAnhanguera School of Bauru, SPDepartment of Biodiversity and Biostatistics Institute of Biosciences São Paulo State University (UNESP), SPDepartment of Infectology Dermatology Diagnostic Imaging and Radiotherapy Botucatu Medical School (FMB) São Paulo State University (UNESP), SPDepartment of Biodiversity and Biostatistics Institute of Biosciences São Paulo State University (UNESP), SPUniversidade Estadual Paulista (UNESP)Anhanguera School of BauruTomazini, Karina Andressa [UNESP]Pereira, Beatriz Aparecida Soares [UNESP]Sylvestre, Tatiane Fernandade Souza Cavalcante, Ricardo [UNESP]de Carvalho, Lídia Raquel [UNESP]Mendes, Rinaldo Poncio [UNESP]2023-07-29T13:45:37Z2023-07-29T13:45:37Z2023-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1590/1678-9199-JVATITD-2022-0045Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 29.1678-91991678-9180http://hdl.handle.net/11449/24849710.1590/1678-9199-JVATITD-2022-00452-s2.0-85149807891Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Venomous Animals and Toxins Including Tropical Diseasesinfo:eu-repo/semantics/openAccess2024-08-15T15:23:14Zoai:repositorio.unesp.br:11449/248497Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-15T15:23:14Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
title Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
spellingShingle Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
Tomazini, Karina Andressa [UNESP]
Blood matrices
Paracoccidioidomycosis
Plasma
Serological diagnosis
Serum
Storage
title_short Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
title_full Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
title_fullStr Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
title_full_unstemmed Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
title_sort Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients
author Tomazini, Karina Andressa [UNESP]
author_facet Tomazini, Karina Andressa [UNESP]
Pereira, Beatriz Aparecida Soares [UNESP]
Sylvestre, Tatiane Fernanda
de Souza Cavalcante, Ricardo [UNESP]
de Carvalho, Lídia Raquel [UNESP]
Mendes, Rinaldo Poncio [UNESP]
author_role author
author2 Pereira, Beatriz Aparecida Soares [UNESP]
Sylvestre, Tatiane Fernanda
de Souza Cavalcante, Ricardo [UNESP]
de Carvalho, Lídia Raquel [UNESP]
Mendes, Rinaldo Poncio [UNESP]
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (UNESP)
Anhanguera School of Bauru
dc.contributor.author.fl_str_mv Tomazini, Karina Andressa [UNESP]
Pereira, Beatriz Aparecida Soares [UNESP]
Sylvestre, Tatiane Fernanda
de Souza Cavalcante, Ricardo [UNESP]
de Carvalho, Lídia Raquel [UNESP]
Mendes, Rinaldo Poncio [UNESP]
dc.subject.por.fl_str_mv Blood matrices
Paracoccidioidomycosis
Plasma
Serological diagnosis
Serum
Storage
topic Blood matrices
Paracoccidioidomycosis
Plasma
Serological diagnosis
Serum
Storage
description Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at –80oC for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at –20oC for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at –80oC for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at –20oC.
publishDate 2023
dc.date.none.fl_str_mv 2023-07-29T13:45:37Z
2023-07-29T13:45:37Z
2023-01-01
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/10.1590/1678-9199-JVATITD-2022-0045
Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 29.
1678-9199
1678-9180
http://hdl.handle.net/11449/248497
10.1590/1678-9199-JVATITD-2022-0045
2-s2.0-85149807891
url http://dx.doi.org/10.1590/1678-9199-JVATITD-2022-0045
http://hdl.handle.net/11449/248497
identifier_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases, v. 29.
1678-9199
1678-9180
10.1590/1678-9199-JVATITD-2022-0045
2-s2.0-85149807891
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Venomous Animals and Toxins Including Tropical Diseases
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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_ 1808128176088416256