ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections

Detalhes bibliográficos
Autor(a) principal: Sidoni, Marli
Data de Publicação: 2018
Outros Autores: Nascimento, Hilton Jorge, Miguez, Mariana, da Silva Junior, José Godinho, Mazur, Carlos, Danelli, Maria das Graças Miranda
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Veterinary Medicine
Texto Completo: https://rbmv.org/BJVM/article/view/911
Resumo: The Feline Immunodeficiency Virus (FIV) is a lentivirus that affects cats worldwide. The detection of the virus’ 24 Kda capsid protein (p24) antibody is the most common way to diagnose FIV infections.  In Brazil, the most used commercial assay for FIV diagnosis is a rapid ELISA test based on the lateral flow principle, with high sensitivity and specificity (SNAP®test). However, the high cost of this imported assay undermines not only the diagnosis of this infection but also its epidemiology. The objective of this study was to optimize and validate an indirect ELISA using the protein p24 recombinant antigen (r-p24) with national technology and maintaining sensitivity and specificity comparable to those of available commercial rapid ELISA tests. Twenty-six reference cats’ sera samples (13 negative and 13 positives for FIV), previously analyzed by PCR and the SNAP® test, were used as a reference to optimize the ELISA. Serum samples of 226 cats from private owners and from shelters of the metropolitan area of Rio de Janeiro were used to validate the assay. The sensitivity and specificity of the r-p24 ELISA were 95.4% and 99.4%, respectively. When compared to the SNAP® Combo Test, the accuracy of r-p24 ELISA was 99%, with a Kappa index of 0.96. Our results indicate that the national technology-based ELISA r-p24 has sensibility and specificity values that are comparable to rapid ELISA kits. This test is therefore recommendable, using national technology for FIV routine diagnosis, research, and epidemiological studies.
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spelling ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infectionsELISA r-p24: desenvolvimento e validação de um ensaio imunoenzimático para o diagnóstico de infecções causadas pelo Vírus da Imunodeficiência FelinaFivELISA r-p24felinosvalidaçãoFIVELISA r-p24felinesvalidation.The Feline Immunodeficiency Virus (FIV) is a lentivirus that affects cats worldwide. The detection of the virus’ 24 Kda capsid protein (p24) antibody is the most common way to diagnose FIV infections.  In Brazil, the most used commercial assay for FIV diagnosis is a rapid ELISA test based on the lateral flow principle, with high sensitivity and specificity (SNAP®test). However, the high cost of this imported assay undermines not only the diagnosis of this infection but also its epidemiology. The objective of this study was to optimize and validate an indirect ELISA using the protein p24 recombinant antigen (r-p24) with national technology and maintaining sensitivity and specificity comparable to those of available commercial rapid ELISA tests. Twenty-six reference cats’ sera samples (13 negative and 13 positives for FIV), previously analyzed by PCR and the SNAP® test, were used as a reference to optimize the ELISA. Serum samples of 226 cats from private owners and from shelters of the metropolitan area of Rio de Janeiro were used to validate the assay. The sensitivity and specificity of the r-p24 ELISA were 95.4% and 99.4%, respectively. When compared to the SNAP® Combo Test, the accuracy of r-p24 ELISA was 99%, with a Kappa index of 0.96. Our results indicate that the national technology-based ELISA r-p24 has sensibility and specificity values that are comparable to rapid ELISA kits. This test is therefore recommendable, using national technology for FIV routine diagnosis, research, and epidemiological studies.O Vírus da Imunodeficiência Felina (FIV) é um lentivírus que afeta gatos em todo o mundo. A detecção do anticorpo para proteína do capsídeo viral de 24 kDa (p24) é a forma mais comum para diagnosticar infecções pelo FIV. No Brasil, o ensaio comercial mais utilizado no diagnóstico FIV é um teste rápido de ELISA (SNAP® test), baseado no princípio de fluxo lateral, com alta sensibilidade e especificidade.  No entanto, o alto custo deste ensaio importado inviabiliza não só o diagnóstico da infecção, como também sua epidemiologia. O objetivo deste trabalho foi aperfeiçoar e validar um teste de ELISA indireto, com tecnologia nacional, utilizando o antígeno recombinante da proteína p24 (r-p24), com sensibilidade eespecificidade comparáveis às dos testes rápidos, comercialmente disponíveis. Vinte e seis amostras de soro gatos (13 negativos e 13 positivos para FIV), previamente analisados por PCR e Snap®, foram utilizadas como amostras de referência para aperfeiçoar o ELISA e 226 amostras de soro de gatos, de proprietários privados ou de abrigos da área metropolitana Rio de Janeiro foram utilizadas para validar o ensaio. O ELISAr-p24 apresentou uma sensibilidade de 95,4% e 99,4% de especificidade quando comparadas às do teste SNAP Combo, uma precisão de 99% e um índice Kappa de 0.96. Nossos resultados indicam que o ELISA r-p24 apresentou sensibilidade e especificidade comparáveis às dos testes rápidos, utilizando tecnologia nacional para diagnóstico, pesquisa e estudos epidemiológicos do FIV.Sociedade de Medicina Veterinária do Estado do Rio de Janeiro.2018-05-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionpeer reviewedAvaliado pelos paresapplication/pdfhttps://rbmv.org/BJVM/article/view/91110.29374/2527-2179.bjvm0293Brazilian Journal of Veterinary Medicine; Vol. 39 No. 2 (2017); 83-89Revista Brasileira de Medicina Veterinária; v. 39 n. 2 (2017); 83-892527-21790100-2430reponame:Brazilian Journal of Veterinary Medicineinstname:Sociedade de Medicina Veterinária do Estado do Rio de Janeiro (SOMVERJ)instacron:SBMVenghttps://rbmv.org/BJVM/article/view/911/748Sidoni, MarliNascimento, Hilton JorgeMiguez, Marianada Silva Junior, José GodinhoMazur, CarlosDanelli, Maria das Graças Mirandainfo:eu-repo/semantics/openAccess2021-03-12T18:05:54Zoai:ojs.rbmv.org:article/911Revistahttps://rbmv.org/BJVMONGhttps://rbmv.org/BJVM/oaicontato.rbmv@gmail.com2527-21790100-2430opendoar:2021-03-12T18:05:54Brazilian Journal of Veterinary Medicine - Sociedade de Medicina Veterinária do Estado do Rio de Janeiro (SOMVERJ)false
dc.title.none.fl_str_mv ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
ELISA r-p24: desenvolvimento e validação de um ensaio imunoenzimático para o diagnóstico de infecções causadas pelo Vírus da Imunodeficiência Felina
title ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
spellingShingle ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
Sidoni, Marli
Fiv
ELISA r-p24
felinos
validação
FIV
ELISA r-p24
felines
validation.
title_short ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
title_full ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
title_fullStr ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
title_full_unstemmed ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
title_sort ELISA r-p24: optimization and validation of an enzyme immunoassay for the diagnosis of Feline Immunodeficiency Virus infections
author Sidoni, Marli
author_facet Sidoni, Marli
Nascimento, Hilton Jorge
Miguez, Mariana
da Silva Junior, José Godinho
Mazur, Carlos
Danelli, Maria das Graças Miranda
author_role author
author2 Nascimento, Hilton Jorge
Miguez, Mariana
da Silva Junior, José Godinho
Mazur, Carlos
Danelli, Maria das Graças Miranda
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Sidoni, Marli
Nascimento, Hilton Jorge
Miguez, Mariana
da Silva Junior, José Godinho
Mazur, Carlos
Danelli, Maria das Graças Miranda
dc.subject.por.fl_str_mv Fiv
ELISA r-p24
felinos
validação
FIV
ELISA r-p24
felines
validation.
topic Fiv
ELISA r-p24
felinos
validação
FIV
ELISA r-p24
felines
validation.
description The Feline Immunodeficiency Virus (FIV) is a lentivirus that affects cats worldwide. The detection of the virus’ 24 Kda capsid protein (p24) antibody is the most common way to diagnose FIV infections.  In Brazil, the most used commercial assay for FIV diagnosis is a rapid ELISA test based on the lateral flow principle, with high sensitivity and specificity (SNAP®test). However, the high cost of this imported assay undermines not only the diagnosis of this infection but also its epidemiology. The objective of this study was to optimize and validate an indirect ELISA using the protein p24 recombinant antigen (r-p24) with national technology and maintaining sensitivity and specificity comparable to those of available commercial rapid ELISA tests. Twenty-six reference cats’ sera samples (13 negative and 13 positives for FIV), previously analyzed by PCR and the SNAP® test, were used as a reference to optimize the ELISA. Serum samples of 226 cats from private owners and from shelters of the metropolitan area of Rio de Janeiro were used to validate the assay. The sensitivity and specificity of the r-p24 ELISA were 95.4% and 99.4%, respectively. When compared to the SNAP® Combo Test, the accuracy of r-p24 ELISA was 99%, with a Kappa index of 0.96. Our results indicate that the national technology-based ELISA r-p24 has sensibility and specificity values that are comparable to rapid ELISA kits. This test is therefore recommendable, using national technology for FIV routine diagnosis, research, and epidemiological studies.
publishDate 2018
dc.date.none.fl_str_mv 2018-05-04
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
peer reviewed
Avaliado pelos pares
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://rbmv.org/BJVM/article/view/911
10.29374/2527-2179.bjvm0293
url https://rbmv.org/BJVM/article/view/911
identifier_str_mv 10.29374/2527-2179.bjvm0293
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://rbmv.org/BJVM/article/view/911/748
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.publisher.none.fl_str_mv Sociedade de Medicina Veterinária do Estado do Rio de Janeiro.
publisher.none.fl_str_mv Sociedade de Medicina Veterinária do Estado do Rio de Janeiro.
dc.source.none.fl_str_mv Brazilian Journal of Veterinary Medicine; Vol. 39 No. 2 (2017); 83-89
Revista Brasileira de Medicina Veterinária; v. 39 n. 2 (2017); 83-89
2527-2179
0100-2430
reponame:Brazilian Journal of Veterinary Medicine
instname:Sociedade de Medicina Veterinária do Estado do Rio de Janeiro (SOMVERJ)
instacron:SBMV
instname_str Sociedade de Medicina Veterinária do Estado do Rio de Janeiro (SOMVERJ)
instacron_str SBMV
institution SBMV
reponame_str Brazilian Journal of Veterinary Medicine
collection Brazilian Journal of Veterinary Medicine
repository.name.fl_str_mv Brazilian Journal of Veterinary Medicine - Sociedade de Medicina Veterinária do Estado do Rio de Janeiro (SOMVERJ)
repository.mail.fl_str_mv contato.rbmv@gmail.com
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