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

... viral genome mutations requires separate approaches such as deep sequencing. A high throughput sequencing platform (ATOPlex) that us. 搜索 当前位置： X-MOL学术 › WaterRes. › 论文详情 RT-qPCRandATOPlexsequencingforthesensitivedetectionofSARS-CoV-2RNAforwastewatersurveillance WaterResearch ( IF 13.400 ) PubDate : 2022-05-16 ,DOI: 10.1016/j.watres.2022.118621 WarishAhmed 1 , AaronBivins 2 , SuzanneMetcalfe 1 , WendyJMSmith 1 , RyanZiels 3 , AsjaKorajkic 4 , BrianMcMinn 4 , TysonEGraber 5 , StuartLSimpson 6 Affiliation  CSIROLandandWater,EcosciencesPrecinct,41BoggoRoad,DuttonPark,QLD4102,Australia. DepartmentofCivil&EnvironmentalEngineering,LouisianaStateUniversity,LA,USA. DepartmentofCivilEngineering,UniversityofBritishColumbia,Vancouver,Canada. UnitedStatesEnvironmentalProtectionAgency,26WMartinLutherKingJr.Drive,Cincinnati,OH45268,USA. Children'sHospitalofEasternOntarioResearchInstitute,OttawaK1H8L1,Canada. CSIROLandandWater,LucasHeights,NSW2234,Australia. 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. 更新日期：2022-05-16   点击分享   查看原文 点击收藏 取消收藏 新增笔记 阅读更多本刊最新论文 本刊介绍/投稿指南 全部期刊列表>> 学术期刊 行业资讯 全球导师 X-MOL问答 求职广场 网址导航 关于我们 帮助中心 客服邮箱：[email protected] 官方微信：X-molTeam2 邮编：100098 地址：北京市海淀区知春路56号中航科技大厦          Copyright©2014-2022北京衮雪科技有限公司AllRightsReserved     京ICP备11026495-2号      京公网安备11010802027423号 down wechat bug bug