Development of a Simple, Rapid, Sensitive, High-throughput Luciferase Reporter Based Microneutralization Test for Measurement of Virus Neutralizing Antibodies Following Respiratory Syncytial Virus Vaccination and Infection

Overview

Journal Vaccine

Specialty Allergy & Immunology

Date 2013 Jun 8

PMID 23742994

Citations 26

Authors

Sandra Fuentes

Roberta L Crim

Judy Beeler

Michael N Teng

Hana Golding

Surender Khurana

Affiliations

Soon will be listed here.

Abstract

We have established a new reporter gene-based RSV neutralization test using Renilla luciferase. The RSV-Luciferase Neutralization Test (RSV-Luc-NeuT) is a simple, rapid, high throughput, and less labor intensive functional serological assay than the traditional RSV-PRNT, capable of measuring a broad range of anti-RSV neutralizing antibodies targeting both RSV-F and RSV-G proteins. Specificity and sensitivity of the RSV-Luc-NeuT are comparable to the RSV-PRNT. Panels of pre-vaccination and post-vaccination animal sera, monoclonal antibodies and animal polyclonal anti-RSV sera confirmed assay specificity. A panel of 60 human sera demonstrated high assay sensitivity for measurement of RSV neutralizing antibodies that strongly correlated with the RSV-PRNT titers (R(2)=0.864). Neutralization in the presence of guinea pig complement (GPC) increased PRNT titers more than the RSV-Luc-NeuT neutralizing antibody titers for these human sera. This newly developed simple, high throughput, RSV-Luc-NeuT could be easily automated and applied in measurement of RSV neutralization titers in large vaccine trials.

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