Quantification and Differentiation of SARS-CoV‑2 Variants in Wastewater for Surveillance

作     者：Teresa Kumblathan Yanming Liu Xiaoli Pang Steve E.Hrudey XChris Le Xing-Fang Li 

作者机构：Division of Analytical and Environmental ToxicologyDepartment of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaCanada T6G 2G3 Division of Diagnostic and Applied MicrobiologyDepartment of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaCanada T6G 2B7 Public Health LaboratoryAlberta Precision LaboratoriesEdmontonAlbertaCanada T6G 2J2 

出 版 物：《Environment & Health》 (环境与健康(英文))

年 卷 期：2023年第1卷第3期

页      面：203-213页

学科分类：083002[工学-环境工程] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 08[工学] 

基　　金：Faculty of Medicine and Dentistry Provincial Laboratory for Public Health, Alberta Precision Laboratories EPCOR New Frontiers in Research Fund Social Sciences and Humanities Research Council of Canada, SSHRC Natural Sciences and Engineering Research Council of Canada, NSERC Canada Research Chairs Alberta Innovates, AI Canadian Institutes of Health Research, CIHR Li Ka Shing Institute of Virology, University of Alberta, LKSIV 

主　　题：COVID-19 SARS-CoV-2 variants of concern Omicron subvariants multiplex RT-qPCR wastewater surveillance 

摘      要：Wastewater surveillance plays an important role in the monitoring of infections of SARS-CoV-2 at the community *** report here the determination of SARS-CoV-2 and differentiation of its variants of concern in 294 wastewater samples collected from two major Canadian cities from May 2021 to March *** overall method of analysis involved extraction of the virus and viral components using electronegative membranes,in situ stabilization and concentration of the viral RNA onto magnetic beads,and direct analysis of the viral RNA on the magnetic *** reverse transcription quantitative polymerase chain reaction(RT-qPCR)assays,targeting specific and naturally selected mutations in SARS-CoV-2,enabled detection and differentiation of the Alpha,Beta,Gamma,Delta,and Omicron *** Omicron triplex RT-qPCR assay targeting three mutations,HV 69−70 deletion,K417N,and L452R,was able to detect and differentiate the Omicron BA.1/BA.3,BA.2/XBB,and BA.4/*** assay had efficiencies of 90−104%for all three mutation targets and a limit of detection of 28 RNA copies per *** of 294 wastewater samples collected over a two-year span showed the concentrations and trends of Alpha,Beta,Gamma,Delta,and Omicron variants as they emerge in two major Canadian cities participating in the wastewater surveillance *** trends of specific variants were consistent with clinical reports for the same *** the beginning of each wave,the corresponding variants were detectable in *** example,RNA concentrations of the BA.2 variant were as high as 10^(4)copies per 100 mL of wastewater collected in January 2022,when approximately only 50−60 clinical cases of BA.2 infection were reported in *** results show that the strategy and highly sensitive assays for the variants of concern in wastewater are potentially useful for the detection of newly emerging SARS-CoV-2 variants and other viruses for future community biomonitoring.