Detection and quantification of the human‐specific HF183 Bacteroides 16S rRNA genetic marker with real‐time PCR for assessment of human faecal pollution in freshwater

摘要: The human-specific HF183 Bacteriodes 16S rRNA genetic marker can be used to detect human faecal pollution in water environments. However, there is currently no method quantify the prevalence of this environmental samples. We developed a real-time polymerase chain reaction (PCR) assay using SYBR Green I detection and To decrease amplicon length suitable size for PCR detection, new reverse primer was designed validated on animal use newly combination with did not specificity but melting curve analysis must always included. This more sensitive than conventional highly reproducible coefficient variation less 1% within an 3% between assays. As Bacteroides species that carries has never been isolated, bacteria determine efficiency. estimated efficiency freshwater ranged from 78% 91% true value average 83+/-4% value. Using simple filtration method, limit quantification 4.7+/-0.3x10(5) markers per litre freshwater. aerobic incubation up 24 days at 4 12 degrees C, 8 28 indicated persisted end period all temperatures.