RT-qPCR and ATOPlex sequencing for the sensitive detection ...

A high throughput sequencing platform (ATOPlex) that uses a multiplex tiled PCR-based enrichment technique has shown promise in detecting variants of ... Conteúdoprincipal1 Busca2 Rodapé3 +A A -A Altocontraste Home Pesquisa RT-qPCRandATOPlexsequencingforthesensitivedetectionofSARS-CoV-2RNAforwastewate RT-qPCRandATOPlexsequencingforthesensitivedetectionofSARS-CoV-2RNAforwastewatersurveillance. Ahmed,Warish; Bivins,Aaron; Metcalfe,Suzanne; Smith,WendyJM; Ziels,Ryan; Korajkic,Asja; McMinn,Brian; Graber,TysonE; Simpson,StuartL. AhmedW;CSIROLandandWater,EcosciencesPrecinct,41BoggoRoad,DuttonPark,QLD4102,Australia.Electronicaddress:[email protected]. BivinsA;DepartmentofCivil&EnvironmentalEngineering,LouisianaStateUniversity,LA,USA. MetcalfeS;CSIROLandandWater,EcosciencesPrecinct,41BoggoRoad,DuttonPark,QLD4102,Australia. SmithWJM;CSIROLandandWater,EcosciencesPrecinct,41BoggoRoad,DuttonPark,QLD4102,Australia. ZielsR;DepartmentofCivilEngineering,UniversityofBritishColumbia,Vancouver,Canada. KorajkicA;UnitedStatesEnvironmentalProtectionAgency,26WMartinLutherKingJr.Drive,Cincinnati,OH45268,USA. McMinnB;UnitedStatesEnvironmentalProtectionAgency,26WMartinLutherKingJr.Drive,Cincinnati,OH45268,USA. GraberTE;Children'sHospitalofEasternOntarioResearchInstitute,OttawaK1H8L1,Canada. SimpsonSL;CSIROLandandWater,LucasHeights,NSW2234,Australia. WaterRes ;220:118621,2022Jul15. Artigo emInglês |MEDLINE |ID:covidwho-1852231 ABSTRACT ABSTRACT Duringthecoronavirusdisease2019(COVID-19)pandemic,wastewatersurveillancehasbecomeanimportanttoolformonitoringthespreadofsevereacuterespiratorysyndromecoronavirus2(SARS-CoV-2)withincommunities.Inparticular,reversetranscription-quantitativePCR(RT-qPCR)hasbeenusedtodetectandquantifySARS-CoV-2RNAinwastewater,whilemonitoringviralgenomemutationsrequiresseparateapproachessuchasdeepsequencing.Ahighthroughputsequencingplatform(ATOPlex)thatusesamultiplextiledPCR-basedenrichmenttechniquehasshownpromiseindetectingvariantsofconcern(VOC)whilealsoprovidingvirusquantitationdata.However,detectionsensitivitiesofbothRT-qPCRandsequencingcanbeimpactedthroughlossesoccurringduringsamplehandling,virusconcentration,nucleicacidextraction,andRT-qPCR.Therefore,processlimitofdetection(PLOD)assessmentsarerequiredtoestimatethegenecopiesoftargetmoleculetoattainspecificprobabilityofdetection.Inthisstudy,wecomparethePLODoffourRT-qPCRassays(USCDCN1andN2,ChinaCDCNandORF1ab)fordetectionofSARS-CoV-2tothatofATOPlexsequencingbyseedingknownconcentrationsofgamma-irradiatedSARS-CoV-2intowastewater.ResultssuggestthatamongtheRT-qPCRassays,USCDCN1wasthemostsensitive,especiallyatlowerSARS-CoV-2seedlevels.However,whenresultsfromallRT-qPCRassayswerecombined,itresultedingreaterdetectionratesthanindividualassays,suggestingthatapplicationofmultipleassaysisbettersuitedforthetracedetectionofSARS-CoV-2fromwastewatersamples.Furthermore,whileATOPlexoffersapromisingapproachtoSARS-CoV-2wastewatersurveillance,thisapproachappearstobelesssensitivecomparedtoRT-qPCRundertheexperimentalconditionsofthisstudy,andmayrequirefurtherrefinements.Nonetheless,thecombinationofRT-qPCRandATOPlexmaybeapowerfultooltosimultaneouslydetect/quantifySARS-CoV-2RNAandmonitoremergingVOCinwastewatersamples. Assuntos COVID-19;SARS-CoV-2;Humanos;RNAViral/genética;TranscriçãoReversa;SARS-CoV-2/genética;ÁguasResiduárias/análise;VigilânciaEpidemiológicaBaseadaemÁguasResiduárias Palavras-chave COVID-19;Concentrationmethod;Detectionlimit;Envelopedvirus;Recovery;SARS-CoV-2;Wastewater Textocompleto https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9109001 https://dx.doi.org/10.1016/j.watres.2022.118621 Imprimir XML PubMedLinks PubMedLinks × BuscarnoGoogle Textocompleto: Disponível Coleções: Basesdedadosinternacionais Basededados: MEDLINE Assuntoprincipal: SARS-CoV-2 / COVID-19 Tipodeestudo: Estudodiagnóstico / Estudoderastreamento Tópicos: Variantes Limite: Humanos Idioma: Inglês Revista: WaterRes Anodepublicação: 2022 Tipodedocumento: Artigo Similares MEDLINE ... LILACS LIS Textocompleto https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9109001 https://dx.doi.org/10.1016/j.watres.2022.118621 Imprimir XML PubMedLinks PubMedLinks × BuscarnoGoogle Textocompleto: Disponível Coleções: Basesdedadosinternacionais Basededados: MEDLINE Assuntoprincipal: SARS-CoV-2 / COVID-19 Tipodeestudo: Estudodiagnóstico / Estudoderastreamento Tópicos: Variantes Limite: Humanos Idioma: Inglês Revista: WaterRes Anodepublicação: 2022 Tipodedocumento: Artigo ConsultaDetalhada × (instance:"regional")AND(year_cluster:("2002")ANDpais_afiliacao:("^iUnitedStates^eEstados"))(instance:"regional")AND(year_cluster:("2002")ANDpais_afiliacao:("^iUnitedStates^eEstados"))(instance:"regional")AND(year_cluster:("2002")ANDpais_afiliacao:("^iUnitedStates^eEstados"))(instance:"regional")AND(year_cluster:("2002")ANDpais_afiliacao:("^iUnitedStates^eEstados")) Fechar Email × Seunome Seuemail Enviarpara Assunto Comentários Exportar × Formatodeexportação: RIS(ReferenceManager,ProCite,EndNote,etc) CSV(Excel,etc) Citação RSS × VisualizarRSS AdicionarRSSnaMinhaBVS Imprimir × Estapágina Referênciasselecionadas(0)