Production of bunyavirus pseudotyped virus v1

Several Bunyavirales families have been listed as being of high pandemic or epidemic risk by the WHO R&D Blueprint. To supportpandemic preparedness and the 100 Days Mission, along with rapid provision of vaccines and therapeutics, the development of tools to assess the immune response is required. Pseudotyped viruses (PV) have been shown to be a suitablealternative to authentic infectious virus to measure virus neutralising activity, a key component of the immune response. They alleviate the need to acquire and amplify viral isolates and do not require high containment facilities. Generating PV of some families within the class Bunyaviricetes is challenging because of a lack of co-localisation of viral glycoproteins atthe vector budding site. This protocol describes a versatile plug-and-play system which has been successfully used for the pseudotyping of two prototype viruses for the family Phenuiviridae, Rift Valley fever virus (RVFV) and for the family Nairoviridae, Crimean-Congo haemorrhagic fever virus (CCHFV). The system was first described by Dr Michael A. Whitt in 2010 (MA Whitt, J Virol Methods 2010 Nov; 169(2): 365-374). The following protocol describes the generation of rVSV*Luc expressing a bunyavirus (e.g. RVFV, CCHFV) glycoprotein. The pseudotyped rVSV*Luc virus can be used to test early steps in virus entry or for the quantification of neutralising antibodies in serological assays.