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A Cell-Based Capture Assay for Rapid Virus Detection
Routine methods for virus detection in clinical specimens rely on a variety of sensitive methods, such as genetic, cell culture and immuno-based assays. It is imperative that the detection assays would be reliable, reproducible, sensitive and rapid. Isolation of viruses from clinical samples is cruc...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602404/ https://www.ncbi.nlm.nih.gov/pubmed/33076296 http://dx.doi.org/10.3390/v12101165 |
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author | Milrot, Elad Makdasi, Efi Politi, Boaz Israely, Tomer Laskar, Orly |
author_facet | Milrot, Elad Makdasi, Efi Politi, Boaz Israely, Tomer Laskar, Orly |
author_sort | Milrot, Elad |
collection | PubMed |
description | Routine methods for virus detection in clinical specimens rely on a variety of sensitive methods, such as genetic, cell culture and immuno-based assays. It is imperative that the detection assays would be reliable, reproducible, sensitive and rapid. Isolation of viruses from clinical samples is crucial for deeper virus identification and analysis. Here we introduce a rapid cell-based assay for isolation and detection of viruses. As a proof of concept several model viruses including West Nile Virus (WNV), Modified Vaccinia Ankara (MVA) and Adenovirus were chosen. Suspended Vero cells were employed to capture the viruses following specific antibody labeling which enables their detection by flow cytometry and immuno-fluorescence microscopy assays. Using flow cytometry, a dose response analysis was performed in which 3.6e4 pfu/mL and 1e6 pfu/mL of MVA and WNV could be detected within two hours, respectively. When spiked to commercial pooled human serum, detection sensitivity was slightly reduced to 3e6 pfu/mL for WNV, but remained essentially the same for MVA. In conclusion, the study demonstrates a robust and rapid methodology for virus detection using flow cytometry and fluorescence microscopy. We propose that this proof of concept may prove useful in identifying future pathogens. |
format | Online Article Text |
id | pubmed-7602404 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76024042020-11-01 A Cell-Based Capture Assay for Rapid Virus Detection Milrot, Elad Makdasi, Efi Politi, Boaz Israely, Tomer Laskar, Orly Viruses Article Routine methods for virus detection in clinical specimens rely on a variety of sensitive methods, such as genetic, cell culture and immuno-based assays. It is imperative that the detection assays would be reliable, reproducible, sensitive and rapid. Isolation of viruses from clinical samples is crucial for deeper virus identification and analysis. Here we introduce a rapid cell-based assay for isolation and detection of viruses. As a proof of concept several model viruses including West Nile Virus (WNV), Modified Vaccinia Ankara (MVA) and Adenovirus were chosen. Suspended Vero cells were employed to capture the viruses following specific antibody labeling which enables their detection by flow cytometry and immuno-fluorescence microscopy assays. Using flow cytometry, a dose response analysis was performed in which 3.6e4 pfu/mL and 1e6 pfu/mL of MVA and WNV could be detected within two hours, respectively. When spiked to commercial pooled human serum, detection sensitivity was slightly reduced to 3e6 pfu/mL for WNV, but remained essentially the same for MVA. In conclusion, the study demonstrates a robust and rapid methodology for virus detection using flow cytometry and fluorescence microscopy. We propose that this proof of concept may prove useful in identifying future pathogens. MDPI 2020-10-15 /pmc/articles/PMC7602404/ /pubmed/33076296 http://dx.doi.org/10.3390/v12101165 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Milrot, Elad Makdasi, Efi Politi, Boaz Israely, Tomer Laskar, Orly A Cell-Based Capture Assay for Rapid Virus Detection |
title | A Cell-Based Capture Assay for Rapid Virus Detection |
title_full | A Cell-Based Capture Assay for Rapid Virus Detection |
title_fullStr | A Cell-Based Capture Assay for Rapid Virus Detection |
title_full_unstemmed | A Cell-Based Capture Assay for Rapid Virus Detection |
title_short | A Cell-Based Capture Assay for Rapid Virus Detection |
title_sort | cell-based capture assay for rapid virus detection |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602404/ https://www.ncbi.nlm.nih.gov/pubmed/33076296 http://dx.doi.org/10.3390/v12101165 |
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