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Discrimination and surveillance of infectious severe acute respiratory syndrome Coronavirus 2 in wastewater using cell culture and RT-qPCR

2022Artigo científico Acesso aberto
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dc.contributor.authorMonteiro, Silvia
dc.contributor.authorRente, Daniela
dc.contributor.authorCunha, Mónica V.
dc.contributor.authorMarques, Tiago A.
dc.contributor.authorCardoso, Eugénia
dc.contributor.authorVilaça, João
dc.contributor.authorCoelho, Norberta
dc.contributor.authorBrôco, Nuno
dc.contributor.authorCarvalho, Marta
dc.contributor.authorSantos, Ricardo
dc.date.accessioned2022-07-01T16:58:20Z
dc.date.available2022-07-01T16:58:20Z
dc.date.issued2022
dc.description.abstractSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA has been extensively detected in raw wastewater in studies exploring wastewater-based epidemiology (WBE) for early warning purposes. Nonetheless, only a few limited studies investigated the presence of SARS-CoV-2 in treated wastewaters to determine the potential health risks across the water cycle. The detection of SARS-CoV-2 has been done mostly by RT-qPCR and ddPCR, which only provides information on the presence of nucleic acids rather than information on potential infectivity. In this study, we set to develop and evaluate the use of viability RT-qPCR for the selective discrimination and surveillance of infectious SARS-CoV-2 in secondary-treated wastewater. Enzymatic (nuclease) and viability dye (Reagent D) pretreatments were applied to infer infectivity through RT-qPCR using porcine epidemic diarrhea virus (PEDV) as a CoV surrogate. Infectivity tests were first performed on PEDV purified RNA, then on infectious and heat-inactivated PEDV, and finally on heat inactivated PEDV spiked in concentrated secondary-treated wastewater. The two viability RT-qPCR methods were then applied to 27 secondary-treated wastewater samples positive for SARS-CoV-2 RNA at the outlet of five large urban wastewater treatment plants in Portugal. Reagent D pretreatment showed similar behavior to cell culture for heat-inactivated PEDV and both viability RT-qPCR methods performed comparably to VERO E6 cell culture for SARS-CoV-2 present in secondary-treated wastewater, eliminating completely the RT-qPCR signal. Our study demonstrated the lack of infectious SARS-CoV-2 viral particles on secondary-treated wastewater through the application of two pretreatment methods for the rapid inference of infectivity through RT-qPCR, showing their potential application in environmental screening. This study addressed a knowledge gap on the public health risks of SARS-CoV-2 across the water cycle.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationSilvia Monteiro, Daniela Rente, Mónica V. Cunha, Tiago A. Marques, Eugénia Cardoso, João Vilaça, Norberta Coelho, Nuno Brôco, Marta Carvalho, Ricardo Santos, Discrimination and surveillance of infectious severe acute respiratory syndrome Coronavirus 2 in wastewater using cell culture and RT-qPCR, Science of The Total Environment, Volume 815, 2022, 152914, ISSN 0048-9697, https://doi.org/10.1016/j.scitotenv.2022.152914.pt_PT
dc.identifier.doi10.1016/j.scitotenv.2022.152914pt_PT
dc.identifier.urihttp://hdl.handle.net/10451/53612
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationCOMPETE (Programa Operacional Competitividade e Internacionalização & Programa Operacional Regional de Lisboa), Portugal 2020 and FEDER funds, in the scope of project “COVIDETECT: Deteção, quantificação e modelação de SARS-CoV-2 em Águas residuais como ferramenta de alerta precoce para a disseminação do vírus na comunidade (Ref. 048467, Aviso N.° 15/SI/2020)pt_PT
dc.relationFCT UIDB/00329/2020pt_PT
dc.relationFCT UIDB/04046/2020pt_PT
dc.relationFCT UIDB/00006/2020pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt_PT
dc.titleDiscrimination and surveillance of infectious severe acute respiratory syndrome Coronavirus 2 in wastewater using cell culture and RT-qPCRpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.startPage152914pt_PT
oaire.citation.titleScience of The Total Environmentpt_PT
oaire.citation.volume815pt_PT
person.familyNameCunha
person.givenNameMonica V.
person.identifier348477
person.identifier.ciencia-id5616-E715-2F31
person.identifier.orcid0000-0003-0401-0276
person.identifier.ridD-5544-2011
person.identifier.scopus-author-id7102711216
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationa1b365bb-7421-4bf5-9cde-62b74c170e69
relation.isAuthorOfPublication.latestForDiscoverya1b365bb-7421-4bf5-9cde-62b74c170e69

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