Detection of mpox virus in wastewater provides forewarning of clinical cases in Canadian cities.
Edgard M Mejia, Nikho A Hizon, Codey E Dueck, Ravinder Lidder, Jade Daigle, Quinn Wonitowy, Nestor G Medina, Umar P Mohammed, Graham W Cox, David Safronetz, Mable Hagan, Jim Strong, Anil Nichani, Michael R Mulvey, Chand S Mangat
Author Information
Edgard M Mejia: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada. Electronic address: edgard.mejia@phac-aspc.gc.ca.
Nikho A Hizon: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Codey E Dueck: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Ravinder Lidder: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Jade Daigle: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Quinn Wonitowy: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Nestor G Medina: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Umar P Mohammed: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Graham W Cox: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
David Safronetz: Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Department of Medical Microbiology and Infectious Diseases, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Mable Hagan: Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Jim Strong: Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Department of Medical Microbiology and Infectious Diseases, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Anil Nichani: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Michael R Mulvey: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Antimicrobial Resistance Nosocomial Infections, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Department of Medical Microbiology and Infectious Diseases, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Chand S Mangat: Wastewater Surveillance Unit, Bacterial Pathogens, AMR, and Wastewater, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Antimicrobial Resistance Nosocomial Infections, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada; Department of Medical Microbiology and Infectious Diseases, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Wastewater-based surveillance (WBS) has shown to be an effective tool in monitoring the spread of SARS-CoV-2 and has helped guide public health actions. Consequently, WBS has expanded to now include the monitoring of mpox virus (MPXV) to contribute to its mitigation efforts. In this study, we demonstrate a unique sample processing and a molecular diagnostic strategy for MPXV detection that can inform on the epidemiological situation of mpox outbreaks through WBS. We conducted WBS for MPXV in 22 Canadian wastewater treatment plants (WWTPs) for 14 weeks. Three MPXV qPCR assays were assessed in this study for the detection of MPXV which include the G2R assays (G2R_WA and G2R_G) developed by the Centers for Disease Control and Prevention (CDC) in 2010, and an in-house-developed assay that we have termed G2R_NML. The G2R_NML assay was designed using reference genomes from the 2022 MPXV outbreak and provides a larger qPCR amplicon size to facilitate Sanger sequencing. Results show that all three assays have similar limits of detection and are able to detect the presence of MPXV in wastewater. The G2R_NML assay produced a significantly greater number of Sanger sequence-confirmed MPXV results compared to the CDC G2R assays. Detection of MPXV was possible where provincial surveillance indicated overall low caseloads, and in some sites forewarning of up to several weeks was observed. Overall, this study proposes that WBS of MPXV provides additional information to help fill knowledge gaps in clinical case-surveillance and is potentially an essential component to the management of mpox.
