An efficient method to enhance recovery and detection of SARS-CoV-2 RNA in wastewater  

作　　者：Teresa Kumblathan Yanming Liu Yuanyuan Qiu Lilly Pang Steve E.Hrudey X.Chris Le Xing-Fang Li 

机构地区：[1]Division of Analytical and Environmental Toxicology,Department of Laboratory Medicine and Pathology,Faculty of Medicine and Dentistry,University of Alberta,Edmonton,Alberta,T6G 2G3,Canada [2]Division of Diagnostic and Applied Microbiology,Department of Laboratory Medicine and Pathology,Faculty of Medicine and Dentistry,University of Alberta,Edmonton,T6G 2G3,Canada [3]Public Health Laboratory,Alberta Precision Laboratories,Edmonton,Alberta,T6G 2G3,Canada

出　　处：《Journal of Environmental Sciences》2023年第8期139-148,共10页环境科学学报（英文版）

基　　金：supported by the Natural Sciences and Engineering Research Council of Canada,the Canada Research Chairs Program,Alberta Innovates,and Alberta Health。

摘　　要：Wastewater surveillance(WS)of SARS-CoV-2 currently requires multiple steps and suffers low recoveries and poor sensitivity.Here,we report an improved analytical method with high sensitivity and recovery to quantify SARS-CoV-2 RNA in wastewater.To improve the recovery,we concentrated SARS-CoV-2 viral particles and RNA from both the solid and aqueous phases of wastewater using an electronegative membrane(EM).The captured viral particles and RNA on the EM were incubated in our newly developed viral inactivation and RNA preservation(VIP)buffer.Subsequently,the RNA was concentrated on magnetic beads and inhibitors removed by washing.Without eluting,the RNA on the magnetic beads was directly detected using reverse transcription quantitative polymerase chain reaction(RT-qPCR).Analysis of SARS-CoV-2 pseudovirus(SARS-CoV-2 RNA in a noninfectious viral coat)spiked to wastewater samples showed an improved recovery of 80%.Analysis of 120 wastewater samples collected twice weekly between May 2021 and February 2022 from two wastewater treatment plants showed 100%positive detection,which agreed with the results independently obtained by a provincial public health laboratory.The concentrations of SARS-CoV-2 RNA in these wastewater samples ranged from 2.4×10^(2) to 2.9×10^(6) copies per 100 mL of wastewater.Our method’s capability of detecting trace and diverse concentrations of SARS-CoV-2 in complex wastewater samples is attributed to the enhanced recovery of SARS-CoV-2 RNA and efficient removal of PCR inhibitors.The improved method for the recovery and detection of viral RNA in wastewater is important for wastewater surveillance,complementing clinical diagnostic tests for public health protection.

关 键 词：WASTEWATER VIRUS Viral RNA Environmental monitoring PATHOGENS SURVEILLANCE 

分 类 号：X703[环境科学与工程—环境工程]