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A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species

Zika virus is a Flavivirus, transmitted via Aedes mosquitos, that causes a range of symptoms including Zika congenital syndrome. Zika has posed a challenging situation for health, public and economic sectors of affected countries. To quantitate Zika virus neutralizing antibody titers in serum sample...

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Autores principales: Bohning, Kelly, Sonnberg, Stephanie, Chen, Hui-Ling, Zahralban-Steele, Melissa, Powell, Timothy, Hather, Greg, Patel, Hetal K., Dean, Hansi J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064526/
https://www.ncbi.nlm.nih.gov/pubmed/33891631
http://dx.doi.org/10.1371/journal.pone.0250516
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author Bohning, Kelly
Sonnberg, Stephanie
Chen, Hui-Ling
Zahralban-Steele, Melissa
Powell, Timothy
Hather, Greg
Patel, Hetal K.
Dean, Hansi J.
author_facet Bohning, Kelly
Sonnberg, Stephanie
Chen, Hui-Ling
Zahralban-Steele, Melissa
Powell, Timothy
Hather, Greg
Patel, Hetal K.
Dean, Hansi J.
author_sort Bohning, Kelly
collection PubMed
description Zika virus is a Flavivirus, transmitted via Aedes mosquitos, that causes a range of symptoms including Zika congenital syndrome. Zika has posed a challenging situation for health, public and economic sectors of affected countries. To quantitate Zika virus neutralizing antibody titers in serum samples, we developed a high throughput plate based Zika virus reporter virus particle (RVP) assay that uses an infective, non-replicating particle encoding Zika virus surface proteins and capsid (CprME) and a reporter gene (Renilla luciferase). This is the first characterization of a Zika virus RVP assay in 384-well format using a Dengue replicon Renilla reporter construct. Serially diluted test sera were incubated with RVPs, followed by incubation with Vero cells. RVPs that have not been neutralized by antibodies in the test sera entered the cells and expressed Renilla luciferase. Quantitative measurements of neutralizing activity were determined using a plate-based assay and commercially available substrate. The principle of limiting the infection to a single round increases the precision of the assay measurements. RVP log(10)EC(50) titers correlated closely with titers determined using a plaque reduction neutralization test (PRNT) (R(2)>95%). The plate-based Zika virus RVP assay also demonstrated high levels of precision, reproducibility and throughput. The assay employs identical reagents for human, rhesus macaque and mouse serum matrices. Spiking studies indicated that the assay performs equally well in different species, producing comparable titers irrespective of the serum species. The assay is conducted in 384-well plates and can be automated to simultaneously achieve high throughput and high reproducibility.
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spelling pubmed-80645262021-05-04 A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species Bohning, Kelly Sonnberg, Stephanie Chen, Hui-Ling Zahralban-Steele, Melissa Powell, Timothy Hather, Greg Patel, Hetal K. Dean, Hansi J. PLoS One Research Article Zika virus is a Flavivirus, transmitted via Aedes mosquitos, that causes a range of symptoms including Zika congenital syndrome. Zika has posed a challenging situation for health, public and economic sectors of affected countries. To quantitate Zika virus neutralizing antibody titers in serum samples, we developed a high throughput plate based Zika virus reporter virus particle (RVP) assay that uses an infective, non-replicating particle encoding Zika virus surface proteins and capsid (CprME) and a reporter gene (Renilla luciferase). This is the first characterization of a Zika virus RVP assay in 384-well format using a Dengue replicon Renilla reporter construct. Serially diluted test sera were incubated with RVPs, followed by incubation with Vero cells. RVPs that have not been neutralized by antibodies in the test sera entered the cells and expressed Renilla luciferase. Quantitative measurements of neutralizing activity were determined using a plate-based assay and commercially available substrate. The principle of limiting the infection to a single round increases the precision of the assay measurements. RVP log(10)EC(50) titers correlated closely with titers determined using a plaque reduction neutralization test (PRNT) (R(2)>95%). The plate-based Zika virus RVP assay also demonstrated high levels of precision, reproducibility and throughput. The assay employs identical reagents for human, rhesus macaque and mouse serum matrices. Spiking studies indicated that the assay performs equally well in different species, producing comparable titers irrespective of the serum species. The assay is conducted in 384-well plates and can be automated to simultaneously achieve high throughput and high reproducibility. Public Library of Science 2021-04-23 /pmc/articles/PMC8064526/ /pubmed/33891631 http://dx.doi.org/10.1371/journal.pone.0250516 Text en © 2021 Bohning et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Bohning, Kelly
Sonnberg, Stephanie
Chen, Hui-Ling
Zahralban-Steele, Melissa
Powell, Timothy
Hather, Greg
Patel, Hetal K.
Dean, Hansi J.
A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title_full A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title_fullStr A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title_full_unstemmed A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title_short A high throughput reporter virus particle microneutralization assay for quantitation of Zika virus neutralizing antibodies in multiple species
title_sort high throughput reporter virus particle microneutralization assay for quantitation of zika virus neutralizing antibodies in multiple species
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064526/
https://www.ncbi.nlm.nih.gov/pubmed/33891631
http://dx.doi.org/10.1371/journal.pone.0250516
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