Packaging of Rift Valley fever virus pseudoviruses and establishment of a neutralization assay method

Abstract

Rift Valley fever (RVF) is an acute, febrile zoonotic disease that is caused by the RVF virus (RVFV). RVF is mainly prevalent on the Arabian Peninsula, the African continent, and several islands in the Indian Ocean near southeast Africa. RVFV has been classified by the World Organisation for Animal Health (OIE) as a category A pathogen. To avoid biological safety concerns associated with use of the pathogen in RVFV neutralization assays, the present study investigated and established an RVFV pseudovirus-based neutralization assay. This study used the human immunodeficiency virus (HIV) lentiviral packaging system and RVFV structural proteins to successfully construct RVFV pseudoviruses. Electron microscopy observation and western blotting indicated that the size, structure, and shape of the packaged pseudoviruses were notably similar to those of HIV lentiviral vectors. Infection inhibition assay results showed that an antibody against RVFV inhibited the infective ability of the RVFV pseudoviruses, and an antibody neutralization assay for RVFV detection was then established. This study has successfully established a neutralization assay based on RVFV pseudoviruses and demonstrated that this method can be used to effectively evaluate antibody neutralization.

Development of a pseudovirus-based assay for measuring neutralizing antibodies against Coxsackievirus A10

“…To date, high-throughput NtAb detection based on pseudoviruses (pNT) has been reported and successfully applied for Poliovirus, Rift Valley fever virus, Human papilloma virus, and HIV, demonstrating high sensitivity, objectivity, and biological safety. [16][17][18][19] Our research group has already constructed numerous types of pseudoviruses relevant to HFMD, including pseudoviruses of enterovirus 71 (EV-A71), CV-A16, CV-B3, CV-B5, and CV-A6, which were used in the development of a pNT assay. [20][21][22][23][24] Here, we report the successful construction of a CV-A10 pseudovirus, and development of a CV-A10 pNT to detect the titre of anti-CV-A10 NtAb.…”