Development of Bacteroides 16S rRNA Gene TaqMan-Based Real-Time PCR Assays for Estimation of Total, Human, and Bovine Fecal Pollution in Water

摘要: Bacteroides species are promising indicators for differentiating livestock and human fecal contamination in water because of their high concentration feces potential host specificity. In this study, a real-time PCR assay was designed to target (AllBac) present human, cattle, equine feces. Direct amplification (without DNA extraction) using the AllBac tested on diluted water. Fecal concentrations threshold cycle were linearly correlated, indicating that can be used estimate total amount Real-time assays also bovine-associated (BoBac) human-associated (HuBac) 16S rRNA genes. Assay specificities bovine, swine, canine, samples. The BoBac specific bovine samples (100% true-positive identification; 0% false-positive identification). HuBac had 100% identification, but it 32% rate with cross-amplification swine creek from three different watersheds. Creek did not inhibit PCR, results correlated those Escherichia coli (r2 = 0.85). percentage attributable sources determined each sample by comparing values obtained assay. These suggest without extraction quantify provide preliminary source identification