
Quantifying viruses in wastewater via RT-qPCR provides total genomic data but does not indicate the virus capsid integrity or the potential risk for human infection. Assessing virus capsid integrity in sewage is important for wastewater-based surveillance, since discharged effluent may pose a public health hazard. While integrity assays using cell cultures can provide this information, they require specialised laboratories and expertise. One solution to overcome this limitation is the use of photo-reactive monoazide dyes (e.g., propidium monoazide [PMAxx]) in a capsid integrity-RT-qPCR assay (ci-RT-qPCR). In this study, we tested the efficiency of PMAxx dye at 50 μM and 100 μM concentrations on live and heat-inactivated model viruses commonly detected in wastewater, including adenovirus (AdV), hepatitis A (HAV), influenza A virus (IAV), and norovirus GI (NoV GI). The 100 μM PMAxx dye concentration effectively differentiated live from heat-inactivated viruses for all targets in buffer solution. This method was then applied to wastewater samples (n = 19) for the detection of encapsulated AdV, enterovirus (EV), HAV, IAV, influenza B virus (IBV), NoV GI, NoV GII, and SARS-CoV-2. Samples were negative for AdV, HAV, IAV, and IBV but positive for EV, NoV GI, NoV GII, and SARS-CoV-2. In the PMAxx-treated samples, EV, NoV GI, and NoV GII showed −0.52–1.15, 0.9–1.51, and 0.31–1.69 log reductions in capsid integrity, indicating a high degree of potentially infectious virus in wastewater. In contrast, SARS-CoV-2 was only detected using RT-qPCR but not after PMAxx treatment, indicating the absence of encapsulated and potentially infectious virus. In conclusion, this study demonstrates the utility of PMAxx dyes to evaluate capsid integrity across a diverse range of viruses commonly monitored in wastewater.
waterborne viruses, Wastewater, Microbiology, enteroviruses, Article, Adenoviridae, Capsid, WBE, Enterovirus Infections, Humans, One Health, WBS, human pathogenic viruses, Coloring Agents, Antigens, Viral, viral inactivation, SARS-CoV-2, viability qPCR, Norovirus, Virion, adenovirus, Hepatitis A, QR1-502, capsid integrity, Capsid Proteins, influenza viruses
waterborne viruses, Wastewater, Microbiology, enteroviruses, Article, Adenoviridae, Capsid, WBE, Enterovirus Infections, Humans, One Health, WBS, human pathogenic viruses, Coloring Agents, Antigens, Viral, viral inactivation, SARS-CoV-2, viability qPCR, Norovirus, Virion, adenovirus, Hepatitis A, QR1-502, capsid integrity, Capsid Proteins, influenza viruses
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