Abstract | Using retroviral pseudotype technology we have recently established assays for the characterisation of neutralising antibodies against rabies virus. Neutralisation assays using sera from vaccinated animals and pseudotyped Challenge Virus Standard-11 detected positive and negative samples with 100% specificity and sensitivity, and titres strongly correlated with the Office International des Epizooties FAVN test (R2 = 0.89). We have also studied the cross-neutralising potency of current rabies vaccines against other lyssavirus genotypes. Compared to conventional live virus assays, studying neutralisation with pseudotypes removes the need for high containment facilities and large sample volumes, therefore increasing the number of laboratories that can undertake active surveillance. Further to this we have adapted the assay so that neutralisation tests can be conducted using luciferase, enhanced green fluorescent protein (GFP) or lacZ as the pseudotype reporter gene. These facets combined make our rabies pseudotype assay an attractive option for use in large vaccine campaigns where it would be advantageous to determine serum neutralising titres in situ. The assay will allow a far greater number of laboratories worldwide, who are presently unable to run live virus test, to be more directly involved in front-line vaccine and antiviral therapy trials, especially in resource limited countries where rabies is a major burden. We aim to develop kits for the distribution of these assays that could be selected with a choice of viral pseudotype and reporter gene. |
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