A New Approach for Active Coronavirus Infection Identification by Targeting the Negative RNA Strand- A Replacement for the Current Positive RNA-based qPCR Detection Method

Abstract

This manuscript describes the development of an alternative method to detect active coronavirus infection, in light of the current COVID-19 pandemic caused by the SARS-CoV-2 virus. The pandemic, which was first identified in Wuhan, China in December 2019, has had a significant impact on global health as well as on the economy and daily life in the world. The current positive RNA-based detection systems are unable to discriminate between replicating and non-replicating viruses, complicating decisions related to quarantine and therapeutic interventions. The proposed method targets the negative strand of the virus and has the potential to effectively distinguish between active and inactive infections, which could provide a more accurate means of determining the spread of the virus and guide more effective public health measures during the current pandemic.

Rapid Detection of Active Coronavirus Infection by Lateral Flow Test Strips: A New Approach to Distinguish Replicating Viruses from Non-Replicating Viruses

“…Here, based on our earlier work, we propose and show the effectiveness of a detection system that can directly target negative-sense viral RNA, produced only during replication. In contrast to positive-stranded fragments, these RNAs should not remain after infection has passed, thereby distinguishing replicating virus from non-replicating virus [17][18][19]. We show that the replicating mouse coronavirus, a model for SARS-CoV [20][21][22], continues to produce both its genome, positive-sense RNA (+ssRNA genome), and antigenome, negative-sense RNA (-ssRNA antigenome), throughout the infection process in cells [5,17].…”

“…In contrast to positive-stranded fragments, these RNAs should not remain after infection has passed, thereby distinguishing replicating virus from non-replicating virus [17][18][19]. We show that the replicating mouse coronavirus, a model for SARS-CoV [20][21][22], continues to produce both its genome, positive-sense RNA (+ssRNA genome), and antigenome, negative-sense RNA (-ssRNA antigenome), throughout the infection process in cells [5,17]. However, once replication has abated, we find that only the +ssRNA genome is still detectable from viruses in host celldepleted culture media, indicating that -ssRNA antigenome may serve as a marker for viral replication competency and may thus indicate infectiousness.…”