Assessment of Rapid and Conventional RT-qPCR-Based Systems for Wastewater Surveillance

Abstract

Conventional wastewater surveillance (WS) relies on highly trained personnel, advanced instrumentation, and significant resources, making the development and use of simple, rapid, and sensitive alternative technologies valuable for reducing costs, time, and labor intensity. For the first time, this study investigated the use of two well-developed rapid systems, including the GeneXpert and LuminUltra, in parallel with a conventional WS reference methodology for the assessment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in three cities: Saskatoon, Prince Albert, and North Battleford, Saskatchewan, Canada. RNA extractions from wastewater samples were carried out for the conventional reference and LuminUltra methods, while GeneXpert was used for both raw and concentrated wastewater samples. Bland–Altman plots showed a combination of systematic bias and random error between these real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR)-based systems. Additionally, results indicated the reasonable performance of GeneXpert in viral detection with a sensitivity rate of >98%, as compared to the conventional reference methodology of 100% and LuminUltra with >65%. A Spearman correlation test showed meaningful relationships between the GeneXpert and conventional reference methodology viral level results across all cities, indicating GeneXpert’s reliability for accurate viral detection and disease prevalence determination, specifically in limited-resource communities, with a shorter processing time and cost-effectiveness in analysis.