Use of GFP-expressing influenza viruses for the detection of influenza virus A/H5N1 neutralizing antibodies.

Abstract:

:The hemagglutination inhibition (HI) assay is used most commonly for the detection of antibodies to influenza viruses. However, for the detection of antibodies to avian influenza viruses of the H5N1 subtype either induced by infection or by vaccination, the HI assay is insensitive. Therefore, the virus neutralization (VN) assay has become the method of choice to detect human serum antibodies directed to these viruses. However, this assay requires a second assay for the detection of residual virus replication, which makes it laborious to perform and less suitable for high throughput testing of large numbers of samples. Here we describe an alternative method for the detection of these antibodies, which is based on the use of reporter viruses that express the green fluorescent protein (GFP) upon infection of target cells. GFP-expressing viruses were generated carrying the HA of a variety of antigenically distinct H5N1 influenza viruses. The method proved easy to perform and could be carried out rapidly. Using a panel of antisera raised against H5N1 influenza viruses, the assay based on GFP expressing viruses was compared with the classical virus neutralization assay and the hemagglutination inhibition assay. In general, the results obtained in these assays correlated well. It was concluded that the assay based on the reporter viruses is an attractive alternative for the classical virus neutralization assay and suitable for large sero-epidemiological studies or for the assessment of vaccine efficacy in clinical trials.

journal_name

Vaccine

journal_title

Vaccine

authors

Rimmelzwaan GF,Verburgh RJ,Nieuwkoop NJ,Bestebroer TM,Fouchier RA,Osterhaus AD

doi

10.1016/j.vaccine.2011.02.082

subject

Has Abstract

pub_date

2011-04-18 00:00:00

pages

3424-30

issue

18

eissn

0264-410X

issn

1873-2518

pii

S0264-410X(11)00330-6

journal_volume

29

pub_type

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