An optimized high-throughput fluorescence plate reader-based RSV neutralization assay

Yong-Peng Sun; Wei Zhang; Qin-Jian Zhao; Jian-Li Cao; Lu-Jing Zhang; Jiang-Yan Xiang; Jun-Yu Si; Xue Lin; Li Chen; Zi-Zheng Zheng; Ning-Shao Xia

doi:10.1016/j.jviromet.2018.07.004

An optimized high-throughput fluorescence plate reader-based RSV neutralization assay

J Virol Methods. 2018 Oct:260:34-40. doi: 10.1016/j.jviromet.2018.07.004. Epub 2018 Jul 9.

Authors

Yong-Peng Sun¹, Wei Zhang², Qin-Jian Zhao³, Jian-Li Cao⁴, Lu-Jing Zhang⁵, Jiang-Yan Xiang⁶, Jun-Yu Si⁷, Xue Lin⁸, Li Chen⁹, Zi-Zheng Zheng¹⁰, Ning-Shao Xia¹¹

Affiliations

¹ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 451464997@qq.com.
² State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health School of Life Sciences, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: aaronzhang@stu.xmu.edu.cn.
³ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: zhaoqinjian@xmu.edu.cn.
⁴ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health School of Life Sciences, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: caojianli@stu.xmu.edu.cn.
⁵ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 104534737@qq.com.
⁶ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health School of Life Sciences, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 1767537307@qq.com.
⁷ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health School of Life Sciences, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 21920122202736@stu.xmu.edu.cn.
⁸ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 634991471@qq.com.
⁹ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: 542773483@qq.com.
¹⁰ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: zhengzizheng@xmu.edu.cn.
¹¹ State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, Fujian 361002, PR China; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health School of Life Sciences, Xiamen University, Xiamen, Fujian 361002, PR China. Electronic address: nsxia@xmu.edu.cn.

PMID: 30003925
DOI: 10.1016/j.jviromet.2018.07.004

Abstract

A licensed vaccine for respiratory syncytial virus (RSV) has yet to be developed, and a reliable and repeatable neutralizing assay is indispensable for vaccine development. Here, we demonstrated an optimized high-throughput RSV neutralization assay that utilizes a fluorescence plate reader (reader) as a substitute for flow cytometry to detect fluorescent signals in RSV-A2 mKate-infected cells. Furthermore, this study tested the influence of virus input and infectivity on the neutralizing assay and highlighted critical factors (together with a suggested protocol) for obtaining stable data using this assay.

Keywords: Flow cytometry; Fluorescence plate reader; Neutralization assay; RSV.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Aged
Aged, 80 and over
Animals
Antibodies, Neutralizing / blood*
Antibodies, Neutralizing / immunology
Antibodies, Viral / blood*
Antibodies, Viral / immunology
Child, Preschool
Fluorescence
Healthy Volunteers
Hep G2 Cells
High-Throughput Screening Assays*
Humans
Mice
Middle Aged
Neutralization Tests*
Reproducibility of Results
Respiratory Syncytial Virus Infections / diagnosis*
Respiratory Syncytial Virus, Human / immunology*

Substances

Antibodies, Neutralizing
Antibodies, Viral