You are here:
Performance of PCR-based assays targeting Bacteroidales genetic markers of human fecal pollution in sewage and fecal samples
Citation:
SHANKS, O. C., K. WHITE, C. A. KELTY, M. SIVAGANESAN, J. BLANNON, M. MECKES, M. VARMA, AND R. A. HAUGLAND. Performance of PCR-based assays targeting Bacteroidales genetic markers of human fecal pollution in sewage and fecal samples. ENVIRONMENTAL SCIENCE AND TECHNOLOGY. American Chemical Society, Washington, DC, 44(16):6281-6288, (2010).
Impact/Purpose:
To inform the public.
Description:
There are numerous PCR-based methods available to characterize human fecal pollution in ambient waters. Each assay employs distinct oligonucleotides and many target different genes and microorganisms leading to potential variations in method performance. Laboratory comparisons are needed to determine which assays are best suited for expensive and time consuming fate/transport and epidemiology studies. We report the host distribution of genetic markers from five end-point PCR and 11 quantitative real-time PCR (qPCR) assays reported to be associated with human feces. Each assay was tested against a reference collection of 54 primary influent sewage samples collected from different geographical locations across the United States and 178 fecal DNA extracts from 23 different animal species., Genetic markers were broadly distributed among sewage samples (96.8-100%) and non-target animal sources (specificity levels ranging from 0% to 96%). For qPCR assays, the abundance of each host-associated genetic marker was measured in all reference samples and compared to quantities of Bacteroidales determined by 16S rRNA qPCR. Experiments indicate that human-associated genetic markers are distributed across abroad range of human populations, but show substantial differences in specificity for human feces.
URLs/Downloads:
PERFORMANCE OF PCR-BASED ASSAYS TARGETING BACTEROIDALES GENETIC MARKERS OF HUMAN FECAL POLLUTION IN SEWAGE AND FECAL SAMPLES
ACS Homepage