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Anti-DENV-NS1 monoclonal antibody for the differential histopathological diagnosis of hemorrhagic fever caused by dengue.

Abstract
Dengue is the most prevalent arboviral disease in humans in tropical and subtropical regions, especially in urban areas, and can cause major epidemics. Although a self-limiting illness, it may sometimes have serious hemorrhagic manifestations, and the outcome of dengue hemorrhagic fever has similar clinical manifestations as in other infections, which could result in death. Therefore, autopsy procedures are required under certain circumstances such as in hemorrhagic fevers, sometimes to confirm or to clarify the diagnosis that may have epidemiological consequences. Normally, the Immunohistochemistry Laboratory of the Pathology Center of Adolfo Lutz Institute receives autopsy samples from different hospitals in Sao Paulo State to confirm a previous diagnosis, especially hemorrhagic fever of infectious etiology. For this diagnosis, we have been using a mouse polyclonal antibody to dengue virus that often does not provide a clear conclusion, because of background staining or no relevant immunostaining, which hampers the histopathological analysis. Accordingly, in the present study, anti-DENV-NS1 monoclonal antibody (4H2) was tested to determine its accuracy in immunohistochemical analysis. Twenty-four autopsy cases of hemorrhagic febrile syndrome showing histopathological alterations compatible with dengue disease were studied: twenty cases were confirmed by RT-PCR for DENV-2 and in four by RT-PCR for yellow fever virus. Samples from autopsied cases of deaths caused by other infectious diseases (two meningitis C and two severe acute respiratory syndrome caused by influenza A H1N1) were included as negative control cases. Positive immunostaining for DENV-NS1 was detected in 16/20 (80%) liver samples and 11/15 (73%) spleen samples from autopsied hemorrhagic dengue patients, whereas the polyclonal antibody detected DENV antigens in 12/20 (60%) liver and in 6/15 (40%) spleen samples from the same cases. Positive results were not obtained with liver biopsy samples from yellow fever or Neisseria meningitides and Flu-A cases. 4H2 mAb recognizes the native protein of the four DENV serotypes in infected cells and did not cross-react with native ZIKV- or CHKV-infected cells by immunohistochemical assay, so it is a useful tool for differential histopathological conclusion of acute febrile hemorrhagic deaths.
AuthorsCristina Takami Kanamura, Roxane Maria Fontes Piazza, Silvia D 'Andretta Iglézias, Cinthya Cirqueira Borges, Marcelo Sansone, Juliana Moutinho Polatto, Dayane Lima Teixeira, Miriam Aparecida da Silva, Leticia Barboza Rocha, Juliana Silva Nogueira, Adriana Yurika Maeda, Fernanda Gisele da Silva Vasami
JournalBrazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] (Braz J Microbiol) Vol. 53 Issue 2 Pg. 777-783 (Jun 2022) ISSN: 1678-4405 [Electronic] Brazil
PMID35129818 (Publication Type: Journal Article)
Copyright© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.
Chemical References
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Viral Nonstructural Proteins
Topics
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Brazil
  • Dengue (diagnosis)
  • Dengue Virus
  • Humans
  • Influenza A Virus, H1N1 Subtype
  • Viral Nonstructural Proteins
  • Zika Virus
  • Zika Virus Infection

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