Performance Characteristics of qPCR Assays TargetingHuman- and Ruminant-Associated Bacteroidetes forMicrobial Source Tracking across Sixteen Countries on Six Continents

摘要

Numerous quantitative PCR assays for microbial fecal source tracking (MST) have been developed and evaluated in recent years. Widespread application has been hindered by a lack of knowledge regarding the geographical stability and hence applicability of such methods beyond the regional level. This study assessed the performance of five previously reported quantitative PCR assays targeting human-, cattle-, or ruminant-associated Bacteroidetes populations on 280 human and animal fecal samples from 16 countries across six continents. The tested cattle-associated markers were shown to be ruminant-associated. The quantitative distributions of marker concentrations in target and nontarget samples proved to be essential for the assessment of assay performance and were used to establish a new metric for quantitative source-specificity. In general, this study demonstrates that stable target populations required for marker-based MST occur around the globe. Ruminant-associated marker concentrations were strongly correlated with total intestinal Bacteroidetes populations and with each other, indicating that the detected ruminant-associated populations seem to be part of the intestinal core microbiome of ruminants worldwide. Consequentlytested ruminant-targeted assays appear to be suitable quantitativeMST tools beyond the regional level while the targeted human-associatedpopulations seem to be less prevalent and stable, suggesting potentialfor improvements in human-targeted methods.