Research Insights
Assessing infectivity of genomic RNA from positive-sense RNA viruses
Several methods have been developed to inactivate virus particles. These inactivation methods must be effective and reliable to prevent the accidental spread of viruses. It is crucial to completely inactivate high-risk agents, such as SARS-CoV-2, before they are removed from high-level biocontainment facilities for further handling. However, it is unclear whether common methods of virus inactivation are effective in inactivating positive-sense RNA viruses. Positive-sense RNA viruses contain RNA genomes of positive polarity and include viruses which can cause serious illness in humans and often result in epidemics and pandemics. The genome of a positive-sense RNA directly serves as messenger RNA (mRNA), thus, can be immediately translated by host cells to produce infectious virions. Therefore, if viral RNA is not properly inactivated during inactivation procedures, the intact RNA can potentially cause infection if introduced into permissive cells. This underscores the need for inactivation protocols that ensure complete inactivation of viral RNA to prevent the accidental release of virus particles into the environment.
Validation for the absence of RNA infectivity after virus inactivation is rarely performed, raising significant biosecurity risks associated with positive-sense RNA viruses. Despite the widespread use of inactivating agents in research labs, there are no standardized protocols for validating RNA infectivity. Testing methods for RNA infectivity are cumbersome and require expertise in RNA, cell culture, and transfection techniques. This study will systematically evaluate the efficacy of existing viral RNA inactivation methods, focusing on their ability to render viral RNA non-infectious. We will develop reagents and protocols required for infectivity testing of viral RNA. By employing appropriate controls and rigorous validation techniques, we will establish standardized protocols for the inactivation of positive-sense viruses and their RNA.
Funder: Centers for Disease Control and Prevention
Amount: $846,421
PI: Lok Joshi, College of Veterinary Medicine