Strategy to develop and validate digital droplet PCR methods for global antimicrobial resistance wastewater surveillance

Antimicrobial resistance (AMR) is a top global health threat, causing infections to become difficult to treat and increasing the risk of disease spread. Wastewater is becoming a new player in AMR surveillance, with real-time PCR and digital droplet PCR technologies being useful analytical methods. However, there are no qPCR and ddPCR methods for AMR surveillance that have been validated according to international standard performance criteria. This study proposes a workflow for developing and validating PCR-based methods for harmonized and global AMR surveillance, including specific sequence databases and microbial collections. The proposed study confirms the applicability of these methods in wastewater samples.