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      Nanosensors based on LSPR are able to serologically differentiate dengue from Zika infections

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          Abstract

          The Flaviviridae virus family was named after the Yellow-fever virus, and the latin term flavi means “of golden color”. Dengue, caused by Dengue virus (DENV), is one of the most important infectious diseases worldwide. A sensitive and differential diagnosis is crucial for patient management, especially due to the occurrence of serological cross-reactivity to other co-circulating flaviviruses. This became particularly important with the emergence of Zika virus (ZIKV) in areas were DENV seroprevalence was already high. We developed a sensitive and specific diagnostic test based on gold nanorods (GNR) functionalized with DENV proteins as nanosensors. These were able to detect as little as one picogram of anti-DENV monoclonal antibodies and highly diluted DENV-positive human sera. The nanosensors could differentiate DENV-positive sera from other flavivirus-infected patients, including ZIKV, and were even able to distinguish which DENV serotype infected individual patients. Readouts are obtained in ELISA-plate spectrophotometers without the need of specific devices.

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          Most cited references 56

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          Immune response to dengue virus and prospects for a vaccine.

          Dengue virus (DENV) is a mosquito-borne member of the Flavivirus genus and includes four serotypes (DENV-1, DENV-2, DENV-3, and DENV-4), each of which is capable of causing dengue fever and dengue hemorrhagic fever/dengue shock syndrome. Serious disease can be seen during primary infection but is more frequent following second infection with a serotype different from that of a previous infection. Infection with wild-type DENV induces high-titered neutralizing antibody that can provide long-term immunity to the homotypic virus and can provide short-term immunity (only several months duration) to a heterotypic DENV. The high level of virus replication seen during both secondary infection with a heterotypic virus and during primary DENV infection in late infancy is a direct consequence of antibody-dependent enhancement of replication. This enhanced virus replication is mediated primarily by preexisting, nonneutralizing, or subneutralizing antibodies to the virion surface antigens that enhance access of the virion-antibody complex to FcγR-bearing cells. Vaccines will need to provide long-term protection against each of the four DENV serotypes by inducing neutralizing antibodies, and live, attenuated and various nonliving virus vaccines are in development.
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            Antibodies to envelope glycoprotein of dengue virus during the natural course of infection are predominantly cross-reactive and recognize epitopes containing highly conserved residues at the fusion loop of domain II.

            The antibody response to the envelope (E) glycoprotein of dengue virus (DENV) is known to play a critical role in both protection from and enhancement of disease, especially after primary infection. However, the relative amounts of homologous and heterologous anti-E antibodies and their epitopes remain unclear. In this study, we examined the antibody responses to E protein as well as to precursor membrane (PrM), capsid, and nonstructural protein 1 (NS1) of four serotypes of DENV by Western blot analysis of DENV serotype 2-infected patients with different disease severity and immune status during an outbreak in southern Taiwan in 2002. Based on the early-convalescent-phase sera tested, the rates of antibody responses to PrM and NS1 proteins were significantly higher in patients with secondary infection than in those with primary infection. A blocking experiment and neutralization assay showed that more than 90% of anti-E antibodies after primary infection were cross-reactive and nonneutralizing against heterologous serotypes and that only a minor proportion were type specific, which may account for the type-specific neutralization activity. Moreover, the E-binding activity in sera of 10 patients with primary infection was greatly reduced by amino acid replacements of three fusion loop residues, tryptophan at position 101, leucine at position 107, and phenylalanine at position 108, but not by replacements of those outside the fusion loop of domain II, suggesting that the predominantly cross-reactive anti-E antibodies recognized epitopes involving the highly conserved residues at the fusion loop of domain II. These findings have implications for our understanding of the pathogenesis of dengue and for the future design of subunit vaccine against DENV as well.
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              Interim Guidance for Interpretation of Zika Virus Antibody Test Results.

              Zika virus is a single-stranded RNA virus in the genus Flavivirus and is closely related to dengue, West Nile, Japanese encephalitis, and yellow fever viruses (1,2). Among flaviviruses, Zika and dengue virus share similar symptoms of infection, transmission cycles, and geographic distribution. Diagnostic testing for Zika virus infection can be accomplished using both molecular and serologic methods. For persons with suspected Zika virus disease, a positive real-time reverse transcription-polymerase chain reaction (rRT-PCR) result confirms Zika virus infection, but a negative rRT-PCR result does not exclude infection (3-7). In these cases, immunoglobulin (Ig) M and neutralizing antibody testing can identify additional recent Zika virus infections (6,7). However, Zika virus antibody test results can be difficult to interpret because of cross-reactivity with other flaviviruses, which can preclude identification of the specific infecting virus, especially when the person previously was infected with or vaccinated against a related flavivirus (8). This is important because the results of Zika and dengue virus testing will guide clinical management. Pregnant women with laboratory evidence of Zika virus infection should be evaluated and managed for possible adverse pregnancy outcomes and be reported to the U.S. Zika Pregnancy Registry or the Puerto Rico Zika Active Pregnancy Surveillance System for clinical follow-up (9,10). All patients with clinically suspected dengue should have proper management to reduce the risk for hemorrhage and shock (11). If serologic testing indicates recent flavivirus infection that could be caused by either Zika or dengue virus, patients should be clinically managed for both infections because they might have been infected with either virus.
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                Author and article information

                Contributors
                dafonseca@icb.ufmg.br
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                9 July 2020
                9 July 2020
                2020
                : 10
                Affiliations
                [1 ]ISNI 0000 0001 2181 4888, GRID grid.8430.f, Laboratório de Virologia Básica e Aplicada, Departamento de Microbiologia, Instituto de Ciências Biológicas, , Universidade Federal de Minas Gerais, ; Belo Horizonte, MG Brazil
                [2 ]ISNI 0000 0004 0635 4678, GRID grid.466576.0, Laboratório de Química de Nanoestruturas de Carbono, , Centro de Desenvolvimento da Tecnologia Nuclear-CDTN/CNEN, ; Belo Horizonte, MG Brazil
                [3 ]ISNI 0000 0001 2181 4888, GRID grid.8430.f, Laboratório de Nanomateriais, Departamento de Física, , Universidade Federal de Minas Gerais, ; Belo Horizonte, MG Brazil
                [4 ]ISNI 0000 0000 9688 4664, GRID grid.472872.c, Fundacao Ezequiel Dias, FUNED, ; Belo Horizonte, MG Brazil
                [5 ]ISNI 0000 0004 0615 5265, GRID grid.419029.7, Laboratório de Pesquisa em Virologia, Departamento de Doenças Infecciosas e Parasitárias, , Faculdade de Medicina de São José Do Rio Preto, ; São José do Rio Preto, SP Brazil
                [6 ]ISNI 0000 0001 2181 4888, GRID grid.8430.f, NanoBioMedical Research Group, Departamento de Física, , Universidade Federal de Minas Gerais, ; Belo Horizonte, MG Brazil
                [7 ]ISNI 0000 0001 2181 4888, GRID grid.8430.f, Centro de Tecnologia de Vacinas, , Universidade Federal de Minas Gerais, ; Belo Horizonte, MG Brazil
                [8 ]ISNI 0000 0004 0615 5265, GRID grid.419029.7, Present Address: Laboratório de Pesquisa em Virologia, Departamento de Doenças Infecciosas e Parasitárias, , Faculdade de Medicina de São José Do Rio Preto, ; São José do Rio Preto, SP Brazil
                Article
                68357
                10.1038/s41598-020-68357-9
                7347616
                © The Author(s) 2020

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                Funding
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo-FAPEMIG (http://www.fapesp.br/)
                Funded by: Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (https://fapemig.br/pt/); Financiadora de Inovação e Pesquisa-FINEP (http://www.finep.gov.br/)
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                biosensors, infectious-disease diagnostics

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