Identification of human fecal pollution: detection and quantification of the human specific HF183 Bacteroides 16S rRNA genetic marker with real-time PCR

摘要

A real-time PCR assay for the detection and quantification in fecal and environmental samples of the human specific HF183 Bacteroides 16S rRNA genetic marker previously found by Bernhard & Field (2000a), was developed and proved to be more sensitive than the conventional HF183 PCR. The limit of detection in canal water was 1.94 x 10~2 copies per μ1 DNA extract and the limit of quantification was 5.83 x 10~2 copies per μ1 DNA extract. The intra- and intervariability coefficients of the real-time assay were less than 1% and 3%, respectively. The amount of marker copy number present per g of wet human feces varied from 10~2 to 10~8 between different individuals. The amount of marker copy number present in wastewater treatment plant influent varied from 10~8 to 10~9 per liter. The marker persisted upto 6 days at 28℃. Whereas, the limit of detection was not reached after 24 days of incubation at 4 and 12℃.