{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T05:15:35Z","timestamp":1787030135652,"version":"build-2736575974"},"reference-count":64,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,12,26]],"date-time":"2023-12-26T00:00:00Z","timestamp":1703548800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon Europe research and innovation programme","award":["101057764"],"award-info":[{"award-number":["101057764"]}]},{"name":"UKRI\/HM Government","award":["101057764"],"award-info":[{"award-number":["101057764"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>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 \u03bcM and 100 \u03bcM 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 \u03bcM 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 \u22120.52\u20131.15, 0.9\u20131.51, and 0.31\u20131.69 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.<\/jats:p>","DOI":"10.3390\/v16010040","type":"journal-article","created":{"date-parts":[[2023,12,27]],"date-time":"2023-12-27T05:43:15Z","timestamp":1703655795000},"page":"40","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Use of Capsid Integrity-qPCR for Detecting Viral Capsid Integrity in Wastewater"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7367-4338","authenticated-orcid":false,"given":"Jessica L.","family":"Kevill","sequence":"first","affiliation":[{"name":"School of Environmental and Natural Sciences, Bangor University, Bangor, Gwynedd LL57 2UW, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7068-3228","authenticated-orcid":false,"given":"Kata","family":"Farkas","sequence":"additional","affiliation":[{"name":"School of Environmental and Natural Sciences, Bangor University, Bangor, Gwynedd LL57 2UW, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Ridding","sequence":"additional","affiliation":[{"name":"School of Environmental and Natural Sciences, Bangor University, Bangor, Gwynedd LL57 2UW, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas","family":"Woodhall","sequence":"additional","affiliation":[{"name":"School of Environmental and Natural Sciences, Bangor University, Bangor, Gwynedd LL57 2UW, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shelagh K.","family":"Malham","sequence":"additional","affiliation":[{"name":"School of Ocean Sciences, Bangor University, Menai Bridge, Anglesey LL59 5AB, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1482-4209","authenticated-orcid":false,"given":"Davey L.","family":"Jones","sequence":"additional","affiliation":[{"name":"School of Environmental and Natural Sciences, Bangor University, Bangor, Gwynedd LL57 2UW, UK"},{"name":"Centre for Sustainable Farming Systems, Food Futures Institute, Murdoch University, Murdoch, WA 6150, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"164561","DOI":"10.1016\/j.scitotenv.2023.164561","article-title":"How Has the COVID-19 Pandemic Impacted Wastewater-Based Epidemiology?","volume":"892","author":"Barcellos","year":"2023","journal-title":"Sci. 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