Novel Methodology for the Detection of Enveloped Viruses

Viral infections in humans cause a huge burden in worldwide healthcare that has increased due to the emergence of new pathogenic viruses, such as in the recent Ebola virus (EBOV) outbreaks. Viral particles in body fluids are often at very low levels, making diagnosis difficult. In order to address t...

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Main Authors: Patricia Resa-Infante, Itziar Erkizia, Jon Ander Nieto-Garai, Maier Lorizate, Nuria Izquierdo-Useros, Javier Martinez-Picado
Format: Article
Language:English
Published: MDPI AG 2020-06-01
Series:Proceedings
Subjects:
Online Access:https://www.mdpi.com/2504-3900/50/1/52
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author Patricia Resa-Infante
Itziar Erkizia
Jon Ander Nieto-Garai
Maier Lorizate
Nuria Izquierdo-Useros
Javier Martinez-Picado
author_facet Patricia Resa-Infante
Itziar Erkizia
Jon Ander Nieto-Garai
Maier Lorizate
Nuria Izquierdo-Useros
Javier Martinez-Picado
author_sort Patricia Resa-Infante
collection DOAJ
description Viral infections in humans cause a huge burden in worldwide healthcare that has increased due to the emergence of new pathogenic viruses, such as in the recent Ebola virus (EBOV) outbreaks. Viral particles in body fluids are often at very low levels, making diagnosis difficult. In order to address this problem, we have developed a new detection platform to isolate and detect different enveloped viruses. We have recently identified that sialic acid-binding Ig‑like lectin 1 (Siglec-1/CD169) is one cellular receptor used by EBOV and HIV-1 to enter myeloid cells, key target cells for infection and pathogenesis. For viral uptake, the V-set domain of this myeloid cell receptor recognizes the gangliosides of viral membranes that were dragged during viral budding from the plasma membrane of infected cells. We took advantage of this specific interaction between Siglec‑1 and viral gangliosides to develop a new detection methodology. We have generated a recombinant protein that contains the V-set domain of Siglec-1 fused to the human IgG Fc domain for anchoring in latex beads. These coated beads allow the isolation of viral particles and their measurement by flow cytometry. We have tested its efficacy to detect HIV-1 and EBOV and its specificity by using anti-Siglec‑1 antibodies that prevent the interaction and serve as a negative control. To test the capacity of our method, we used synthetic liposomes to assess the effect of ganglioside concentration in membranes as well as the size of viral particles. This methodology would facilitate the diagnosis of infections by concentrating viral particles in a fast and direct method. At a time when global human mobility facilitates the dissemination of infectious agents, our approach represents a rapid and effective method to maximize the identification of both known and emerging enveloped viruses as part of public health viral surveillance strategies.
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spelling doaj.art-a313109a431544198119d8b1f1c7e79d2023-11-20T03:50:12ZengMDPI AGProceedings2504-39002020-06-015015210.3390/proceedings2020050052Novel Methodology for the Detection of Enveloped VirusesPatricia Resa-Infante0Itziar Erkizia1Jon Ander Nieto-Garai2Maier Lorizate3Nuria Izquierdo-Useros4Javier Martinez-Picado5IrsiCaixa AIDS Research Institute, Ctra. de Canyet s/n, 08916 Badalona, SpainIrsiCaixa AIDS Research Institute, Ctra. de Canyet s/n, 08916 Badalona, SpainInstituto Biofisika (CSIC, UPV/EHU), University of the Basque Country (UPV/EHU), 48940 Bilbao, SpainInstituto Biofisika (CSIC, UPV/EHU) and Department of Biochemistry and Molecular Biology, University of the Basque Country (UPV/EHU), 48940 Bilbao, SpainIrsiCaixa AIDS Research Institute, Ctra. de Canyet s/n, 08916 Badalona, SpainIrsiCaixa AIDS Research Institute, Ctra. de Canyet s/n, 08916 Badalona, SpainViral infections in humans cause a huge burden in worldwide healthcare that has increased due to the emergence of new pathogenic viruses, such as in the recent Ebola virus (EBOV) outbreaks. Viral particles in body fluids are often at very low levels, making diagnosis difficult. In order to address this problem, we have developed a new detection platform to isolate and detect different enveloped viruses. We have recently identified that sialic acid-binding Ig‑like lectin 1 (Siglec-1/CD169) is one cellular receptor used by EBOV and HIV-1 to enter myeloid cells, key target cells for infection and pathogenesis. For viral uptake, the V-set domain of this myeloid cell receptor recognizes the gangliosides of viral membranes that were dragged during viral budding from the plasma membrane of infected cells. We took advantage of this specific interaction between Siglec‑1 and viral gangliosides to develop a new detection methodology. We have generated a recombinant protein that contains the V-set domain of Siglec-1 fused to the human IgG Fc domain for anchoring in latex beads. These coated beads allow the isolation of viral particles and their measurement by flow cytometry. We have tested its efficacy to detect HIV-1 and EBOV and its specificity by using anti-Siglec‑1 antibodies that prevent the interaction and serve as a negative control. To test the capacity of our method, we used synthetic liposomes to assess the effect of ganglioside concentration in membranes as well as the size of viral particles. This methodology would facilitate the diagnosis of infections by concentrating viral particles in a fast and direct method. At a time when global human mobility facilitates the dissemination of infectious agents, our approach represents a rapid and effective method to maximize the identification of both known and emerging enveloped viruses as part of public health viral surveillance strategies.https://www.mdpi.com/2504-3900/50/1/52Siglec-1HIV-1Ebola virusenveloped particlesIsolation methods
spellingShingle Patricia Resa-Infante
Itziar Erkizia
Jon Ander Nieto-Garai
Maier Lorizate
Nuria Izquierdo-Useros
Javier Martinez-Picado
Novel Methodology for the Detection of Enveloped Viruses
Proceedings
Siglec-1
HIV-1
Ebola virus
enveloped particles
Isolation methods
title Novel Methodology for the Detection of Enveloped Viruses
title_full Novel Methodology for the Detection of Enveloped Viruses
title_fullStr Novel Methodology for the Detection of Enveloped Viruses
title_full_unstemmed Novel Methodology for the Detection of Enveloped Viruses
title_short Novel Methodology for the Detection of Enveloped Viruses
title_sort novel methodology for the detection of enveloped viruses
topic Siglec-1
HIV-1
Ebola virus
enveloped particles
Isolation methods
url https://www.mdpi.com/2504-3900/50/1/52
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