Methods for successful inactivation of Rift Valley fever virus in infected mosquitoes.

ABSTRACT

Ensuring the successful inactivation of select agent material is critical for maintaining compliance with federal regulations and safeguarding laboratory personnel from exposure to dangerous pathogens. Rift Valley fever virus (RVFV), naturally transmitted by mosquitoes, is classified as a select agent by the CDC and USDA due to its potential to cause significant economic losses to the livestock industry and its demonstrated potential to emerge into naïve geographic areas. Herein we describe several effective inactivation procedures for RVFV infected mosquito samples. We also demonstrate the vaccine strain MP-12 can be used as an appropriate analog for inactivation testing and describe a method of validating inactivation using Amicon filters. Briefly, we show the following inactivation methods are all effective at inactivating RVFV and MP-12 by following the manufacturers'/established protocols 4 % paraformaldehyde, Trizol LS (ThermoFisher Scientific), MagMAX™-96 Viral RNA Isolation Kit (ThermoFisher Scientific), and Mag-Bind® Viral DNA/RNA 96 Kit (Omega Bio-Tek).