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Development of Rapid Canine Fecal Source Identification PCR-based Assays
Citation:
Green, H. C., Karen M. White, C. A. Kelty, AND O. C. Shanks. Development of Rapid Canine Fecal Source Identification PCR-based Assays. ENVIRONMENTAL SCIENCE & TECHNOLOGY. American Chemical Society, Washington, DC, 48(19):11453-11461, (2014).
Impact/Purpose:
To inform the public on the extent to which dogs contribute to aquatic fecal contamination.
Description:
The extent to which dogs contribute to aquatic fecal contamination is unknown despite the potential for zoonotic transfer of harmful human pathogens. We used Genome Fragment Enrichment (GFE) to identify novel non-ribosomal microbial genetic markers potentially useful for detecting dog fecal contamination with PCR-based methods in environmental samples. Of the 679 sequences obtained from GFE, we used 84 for the development of PCR assays targeting putative canine-associated genetic markers. Twelve genetic markers were shown to be prevalent among dog fecal samples and were rarely found in other animals. Three assays, DG3, DG37, and DG72, performed best in terms of specificity and sensitivity and were used for the development of SYBR® Green and TaqMan® quantitative PCR (qPCR) assays. qPCR analysis of 244 fecal samples collected from a wide geographic range indicated that marker concentrations were below limits of detection in non-canine hosts. In addition, these markers were detected in five out of 18 urban stormwater samples, suggesting that they persist long enough under environmental conditions to be useful for water quality applications. These new assays will be useful to water resource managers who wish to use fecal source identification tools to help manage local water resources impacted by dog feces.
