Viral pathogens have significantly impacted public health and the economy. These viruses can spread through the environment, which is one of the main exposure routes for humans. There are two approaches to monitor the infectivity of viruses in the environment depending on host cells. First, culture-dependent assays use host cells to directly measure the viral infectivity and are regarded as the gold standard. However, many viruses do not have appropriate host cell lines that can be consistently grown in the laboratory. Besides, a diverse population of viruses in environmental samples means different host cells are required for the culture-dependent assays. Second, culture-independent assays quantify the number of viral genomes and multiply it by an assumed ratio of infectivity to the genome copy number to calculate the viral infectivity. However, most of the virus inactivation is actually related to other viral structures such as viral proteins or envelopes in the environment, and thus monitoring the number of viral genomes is not an accurate method.<br/>We propose a novel culture-independent assay that is also capable of estimating the viral infectivity. The key feature of our assay is using magnetic beads coated with the host cell receptors to selectively bind infectious viruses including rotavirus, Tulane virus (monkey virus with the same receptor as human norovirus), adenovirus, transmissible gastroenteritis virus, and NL63 (human coronavirus with the same receptor as SARS-CoV-2). This assay needs only magnets for concentrating the viruses in a few hours while the total genome-based assay requires complex instruments and takes a longer time. More importantly, we confirmed the novel assay explains changes in viral infectivity in wastewater (which was determined by plaque assays) much better than the total genome-based assay. We also found the PGM-MBs show an even lower detection limit than the conventional assay. We vision that the novel assay contributes to better control of infectious viruses in the environment, which ultimately protect the public from infectious viral diseases.

Culture-independent surveillance of infectious viruses in the environment

ACS Spring Meeting 2021

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