Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFMG |
Texto Completo: | https://doi.org/10.1590/0074-02760200287 http://hdl.handle.net/1843/50610 https://orcid.org/0000-0002-3629-8529 https://orcid.org/0000-0002-5253-4261 https://orcid.org/0000-0002-9773-0194 https://orcid.org/0000-0002-5732-7173 https://orcid.org/0000-0002-8026-7017 https://orcid.org/0000-0002-1471-0084 https://orcid.org/0000-0003-1489-794X https://orcid.org/0000-0002-5365-4329 |
Resumo: | BACKGROUND: The heat-labile nature of Dengue virus (DENV) in serum samples must be considered when applying routine diagnostic tests to avoid issues that could impact the accuracy of test results with direct implications for case management and disease reporting. OBJECTIVES: To check if pre-analytical variables, such as storage time and temperature, have an impact on the accuracy of the main routine diagnostic tests for dengue. METHODS: Virus isolation, reverse transcription real-time polymerase chain reaction (RT PCR) and NS1 enzyme-linked immunosorbent assay (ELISA) were evaluated using 84 samples submitted to different pre-analytical conditions. FINDINGS: Sensitivity and negative predictive value were directly affected by sample storage conditions. RT-PCR and virus isolation showed greater dependence on well-conserved samples for an accurate diagnosis. Interestingly, even storage at -30ºC for a relatively short time (15 days) was not adequate for accurate results using virus isolation and RT-PCR tests. On the other hand, NS1 ELISA showed no significant reduction in positivity for aliquots tested under the same conditions as in the previous tests. MAIN CONCLUSIONS: Our results support the stability of the NS1 marker in ELISA diagnosis and indicate that the accuracy of routine tests such as virus isolation and RT-PCR is significantly affected by inadequate transport and storage conditions of serum samples. |
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2023-03-02T17:46:46Z2023-03-02T17:46:46Z2020-11115https://doi.org/10.1590/0074-027602002871678-8060http://hdl.handle.net/1843/50610https://orcid.org/0000-0002-3629-8529https://orcid.org/0000-0002-5253-4261https://orcid.org/0000-0002-9773-0194https://orcid.org/0000-0002-5732-7173https://orcid.org/0000-0002-8026-7017https://orcid.org/0000-0002-1471-0084https://orcid.org/0000-0003-1489-794Xhttps://orcid.org/0000-0002-5365-4329BACKGROUND: The heat-labile nature of Dengue virus (DENV) in serum samples must be considered when applying routine diagnostic tests to avoid issues that could impact the accuracy of test results with direct implications for case management and disease reporting. OBJECTIVES: To check if pre-analytical variables, such as storage time and temperature, have an impact on the accuracy of the main routine diagnostic tests for dengue. METHODS: Virus isolation, reverse transcription real-time polymerase chain reaction (RT PCR) and NS1 enzyme-linked immunosorbent assay (ELISA) were evaluated using 84 samples submitted to different pre-analytical conditions. FINDINGS: Sensitivity and negative predictive value were directly affected by sample storage conditions. RT-PCR and virus isolation showed greater dependence on well-conserved samples for an accurate diagnosis. Interestingly, even storage at -30ºC for a relatively short time (15 days) was not adequate for accurate results using virus isolation and RT-PCR tests. On the other hand, NS1 ELISA showed no significant reduction in positivity for aliquots tested under the same conditions as in the previous tests. MAIN CONCLUSIONS: Our results support the stability of the NS1 marker in ELISA diagnosis and indicate that the accuracy of routine tests such as virus isolation and RT-PCR is significantly affected by inadequate transport and storage conditions of serum samples.JUSTIFICATIVA: A natureza termolábil do vírus da dengue (DENV) em amostras de soro deve ser considerada ao aplicar testes de diagnóstico de rotina para evitar problemas que possam afetar a precisão dos resultados do teste com implicações diretas para o gerenciamento de casos e notificação da doença. OBJETIVOS: Verificar se variáveis pré-analíticas, como tempo de armazenamento e temperatura, interferem na acurácia dos principais testes diagnósticos de rotina para dengue. MÉTODOS: O isolamento viral, a reação em cadeia da polimerase em tempo real com transcrição reversa (RT PCR) e o ensaio imunoenzimático (ELISA) de NS1 foram avaliados em 84 amostras submetidas a diferentes condições pré-analíticas. DADOS: A sensibilidade e o valor preditivo negativo foram diretamente afetados pelas condições de armazenamento das amostras. RT-PCR e isolamento do vírus mostraram maior dependência de amostras bem conservadas para um diagnóstico preciso. Curiosamente, mesmo o armazenamento a -30ºC por um tempo relativamente curto (15 dias) não foi adequado para resultados precisos usando isolamento viral e testes de RT-PCR. Por outro lado, o NS1 ELISA não apresentou redução significativa da positividade para as alíquotas testadas nas mesmas condições dos testes anteriores. CONCLUSÕES PRINCIPAIS: Nossos resultados apóiam a estabilidade do marcador NS1 no diagnóstico ELISA e indicam que a precisão de testes de rotina, como isolamento viral e RT-PCR, é significativamente afetada por condições inadequadas de transporte e armazenamento de amostras de soro.Outra AgênciaengUniversidade Federal de Minas GeraisUFMGBrasilHCL - HOSPITAL DAS CLINICASDengueVírus da dengueTestes laboratoriaisDengueRNASerum samplesStorage conditionsDiagnosisDengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followedDiagnóstico da dengue: é provável que ocorram imprecisões graves se as condições pré-analíticas não forem estritamente seguidasinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttps://memorias.ioc.fiocruz.br/article/10779/0287-dengue-diagnostics-serious-inaccuracies-are-likely-to-occur-if-pre-analytical-conditions-are-not-strictly-followed#backFelipe Campos de Melo IaniMyrian Morato DuarteAna Carolina Barbosa CaetanoJéssica Caroline Wenceslau CocovichFrederico Figueiredo AmâncioMaira Alves PereiraTalita Émile Ribeiro AdelinoSérgio CaldasMarcos Vinícius Ferreira SilvaGlauco de Carvalho Pereiraapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSELicense.txtLicense.txttext/plain; charset=utf-82042https://repositorio.ufmg.br/bitstream/1843/50610/1/License.txtfa505098d172de0bc8864fc1287ffe22MD51ORIGINALDengue diagnostics serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed.pdfDengue diagnostics serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed.pdfapplication/pdf550533https://repositorio.ufmg.br/bitstream/1843/50610/2/Dengue%20diagnostics%20serious%20inaccuracies%20are%20likely%20to%20occur%20if%20pre-analytical%20conditions%20are%20not%20strictly%20followed.pdf5234516f828217a59f7c273e56b20078MD521843/506102023-03-02 14:46:47.097oai:repositorio.ufmg.br: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Repositório de PublicaçõesPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-03-02T17:46:47Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false |
dc.title.pt_BR.fl_str_mv |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
dc.title.alternative.pt_BR.fl_str_mv |
Diagnóstico da dengue: é provável que ocorram imprecisões graves se as condições pré-analíticas não forem estritamente seguidas |
title |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
spellingShingle |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed Felipe Campos de Melo Iani Dengue RNA Serum samples Storage conditions Diagnosis Dengue Vírus da dengue Testes laboratoriais |
title_short |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
title_full |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
title_fullStr |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
title_full_unstemmed |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
title_sort |
Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed |
author |
Felipe Campos de Melo Iani |
author_facet |
Felipe Campos de Melo Iani Myrian Morato Duarte Ana Carolina Barbosa Caetano Jéssica Caroline Wenceslau Cocovich Frederico Figueiredo Amâncio Maira Alves Pereira Talita Émile Ribeiro Adelino Sérgio Caldas Marcos Vinícius Ferreira Silva Glauco de Carvalho Pereira |
author_role |
author |
author2 |
Myrian Morato Duarte Ana Carolina Barbosa Caetano Jéssica Caroline Wenceslau Cocovich Frederico Figueiredo Amâncio Maira Alves Pereira Talita Émile Ribeiro Adelino Sérgio Caldas Marcos Vinícius Ferreira Silva Glauco de Carvalho Pereira |
author2_role |
author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Felipe Campos de Melo Iani Myrian Morato Duarte Ana Carolina Barbosa Caetano Jéssica Caroline Wenceslau Cocovich Frederico Figueiredo Amâncio Maira Alves Pereira Talita Émile Ribeiro Adelino Sérgio Caldas Marcos Vinícius Ferreira Silva Glauco de Carvalho Pereira |
dc.subject.por.fl_str_mv |
Dengue RNA Serum samples Storage conditions Diagnosis |
topic |
Dengue RNA Serum samples Storage conditions Diagnosis Dengue Vírus da dengue Testes laboratoriais |
dc.subject.other.pt_BR.fl_str_mv |
Dengue Vírus da dengue Testes laboratoriais |
description |
BACKGROUND: The heat-labile nature of Dengue virus (DENV) in serum samples must be considered when applying routine diagnostic tests to avoid issues that could impact the accuracy of test results with direct implications for case management and disease reporting. OBJECTIVES: To check if pre-analytical variables, such as storage time and temperature, have an impact on the accuracy of the main routine diagnostic tests for dengue. METHODS: Virus isolation, reverse transcription real-time polymerase chain reaction (RT PCR) and NS1 enzyme-linked immunosorbent assay (ELISA) were evaluated using 84 samples submitted to different pre-analytical conditions. FINDINGS: Sensitivity and negative predictive value were directly affected by sample storage conditions. RT-PCR and virus isolation showed greater dependence on well-conserved samples for an accurate diagnosis. Interestingly, even storage at -30ºC for a relatively short time (15 days) was not adequate for accurate results using virus isolation and RT-PCR tests. On the other hand, NS1 ELISA showed no significant reduction in positivity for aliquots tested under the same conditions as in the previous tests. MAIN CONCLUSIONS: Our results support the stability of the NS1 marker in ELISA diagnosis and indicate that the accuracy of routine tests such as virus isolation and RT-PCR is significantly affected by inadequate transport and storage conditions of serum samples. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-11 |
dc.date.accessioned.fl_str_mv |
2023-03-02T17:46:46Z |
dc.date.available.fl_str_mv |
2023-03-02T17:46:46Z |
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://hdl.handle.net/1843/50610 |
dc.identifier.doi.pt_BR.fl_str_mv |
https://doi.org/10.1590/0074-02760200287 |
dc.identifier.issn.pt_BR.fl_str_mv |
1678-8060 |
dc.identifier.orcid.pt_BR.fl_str_mv |
https://orcid.org/0000-0002-3629-8529 https://orcid.org/0000-0002-5253-4261 https://orcid.org/0000-0002-9773-0194 https://orcid.org/0000-0002-5732-7173 https://orcid.org/0000-0002-8026-7017 https://orcid.org/0000-0002-1471-0084 https://orcid.org/0000-0003-1489-794X https://orcid.org/0000-0002-5365-4329 |
url |
https://doi.org/10.1590/0074-02760200287 http://hdl.handle.net/1843/50610 https://orcid.org/0000-0002-3629-8529 https://orcid.org/0000-0002-5253-4261 https://orcid.org/0000-0002-9773-0194 https://orcid.org/0000-0002-5732-7173 https://orcid.org/0000-0002-8026-7017 https://orcid.org/0000-0002-1471-0084 https://orcid.org/0000-0003-1489-794X https://orcid.org/0000-0002-5365-4329 |
identifier_str_mv |
1678-8060 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
dc.publisher.initials.fl_str_mv |
UFMG |
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Brasil |
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HCL - HOSPITAL DAS CLINICAS |
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Universidade Federal de Minas Gerais |
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