Validation and application of a standardized quantitative PCR assay for the assessment of antimicrobial resistance genes in surface water

This study developed and validated a standardized quantitative PCR (qPCR) method to detect and monitor antimicrobial resistance genes (ARGs) in surface water. The assay targets 47 resistance-related genes and showed very high performance, with 97.7% sensitivity and 98.7% specificity compared with whole-genome sequencing. When applied to surface water samples from several national parks in Alaska, the method detected 19 different resistance genes, with higher diversity and abundance of ARGs at sites with greater human visitation. These findings demonstrate that standardized qPCR approaches can reliably monitor AMR in environmental waters and suggest that human activity may influence the presence of resistance genes even in relatively remote ecosystems, highlighting the importance of environmental surveillance within a One Health AMR monitoring framework.