Seasonal surveillance of various pathogenic viruses in wastewater using a high-throughput PCR system and comparison of two concentration methods

In recent years, the analysis of viruses in wastewater has become a recognized method to monitor the circulation of pathogenic viruses within the populations. This non-invasive and integrated approach is all the more strategic as many viral pathogens can be released in sewage. There is currently no standard method for the virological analysis of wastewater, and various protocols have been developed to concentrate and detect viruses in this matrix. This study aims to develop a high-throughput qPCR detection system to describe the occurrence of 30 human enteric and respiratory viruses in influent and effluent wastewater samples collected in the Paris area during two periods in 2023. The influence of the concentration method (ultracentrifugation or aluminium hydroxide adsorption-precipitation) for the recovery of viruses from wastewater samples was also investigated. The ultracentrifugation-based method allowed a higher viral recovery (4.95-100 % versus 0.77-33 % for the precipitation-based method), especially in effluent waters. The PCR array revealed the presence of many pathogenic viruses, with marked seasonal variations mainly for respiratory viruses. This system is particularly suitable for the rapid, sensitive, and specific detection of multiple viruses in wastewater.