Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed

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. To check if pre-analytical variables, such as st...

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Bibliographic Details
Published in:	Memórias do Instituto Oswaldo Cruz Vol. 115; p. e200287
Main Authors:	Iani, Felipe Campos de Melo, Caetano, Ana Carolina Barbosa, Cocovich, Jéssica Caroline Wenceslau, Amâncio, Frederico Figueiredo, Pereira, Maira Alves, Adelino, Talita Émile Ribeiro, Caldas, Sérgio, Silva, Marcos Vinícius Ferreira, Pereira, Glauco de Carvalho, Duarte, Myrian Morato
Format:	Journal Article
Language:	English
Published:	Brazil Instituto Oswaldo Cruz, Ministério da Saúde 2021 Fundação Oswaldo Cruz (FIOCRUZ)
Subjects:	Antibodies, Viral - blood Antigens, Viral - blood Antigens, Viral - immunology dengue Dengue - blood Dengue - diagnosis Dengue - virology Dengue Virus - genetics Dengue Virus - immunology Dengue Virus - isolation & purification diagnosis Enzyme-Linked Immunosorbent Assay - methods Humans Immunologic Tests - methods Original PARASITOLOGY Predictive Value of Tests Reverse Transcriptase Polymerase Chain Reaction - methods RNA Sensitivity and Specificity serum samples storage conditions TROPICAL MEDICINE Viral Nonstructural Proteins - genetics Viral Nonstructural Proteins - immunology diagnosis storage conditions serum samples RNA dengue
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Summary:	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. 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. 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. 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. 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 AUTHORS’ CONTRIBUTION: FCMI, FFA and GCP conceived and designed the experiments; FCMI, ACBC, JCWC, MAP, TERA, MMD and MVFS performed the experiments; FCMI, SC, MVFS and MMD analysed the data; FCMI and GCP contributed with reagents/materials/analytical tools; FCMI, SC and MMD wrote and/or revised the paper.
ISSN:	0074-0276 1678-8060 1678-8060
DOI:	10.1590/0074-02760200287