The use of equine influenza pseudotypes for serological screening

Scott, Simon D. and Molesti, Eleonora and Temperton, Nigel J. and Ferrara, Francesca and Böttcher-Friebertshäuser, Eva and Daly, Janet (2013) The use of equine influenza pseudotypes for serological screening. In: Society for General Microbiology Spring Meeting, 25-28 Mar 2013, Manchester. (Unpublished) (doi:MA09/03) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Abstract

Standard assays used for influenza serology present certain practical issues, such as inter-laboratory variability, complex protocols and necessity for handling certain virus strains in high biological containment facilities. In an attempt to address this, avian and human influenza HA pseudotyped retroviruses have been successfully employed in antibody neutralisation assays. In this study we have generated equine influenza pseudotyped lentiviruses for serological screening. This was achieved by co-transfection of HEK293T cells with plasmids expressing the haemagglutinin (HA) protein of different H3N8 subtype equine influenza virus strains, HIV gag-pol and firefly luciferase reporter genes and harvesting virus from supernatant. To produce infective pseudotype particles it was also necessary to co-transfect a plasmid encoding the TMPRSS2 endoprotease to cleave HA. High titre pseudotype virus (PV) was then used in PV antibody neutralisation assays (PVNAs) to successfully distinguish between vaccinated and non-vaccinated equines. The sera were also screened by single radial haemolysis (SRH) assay. There was a 65% statistical correlation between the results of the two assays, with the PVNA assay appearing more slightly more sensitive. Future work will extend the testing of the PVNA with a larger cohort of serum samples to assess sensitivity/specificity, inter/intra-laboratory variability and to define a protective titre.

Item Type: Conference or workshop item (Poster)
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 22 Apr 2013 13:17 UTC
Last Modified: 04 Dec 2017 11:33 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/33652 (The current URI for this page, for reference purposes)
Scott, Simon D.: https://orcid.org/0000-0002-8290-0461
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