Development of Rapid Canine Fecal Source Identification PCR-Based Assays

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 nonribosomal microbial genetic markers potentially useful for detecting dog fecal contamina...

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Bibliographic Details
Published in:Environmental science & technology 2014-10, Vol.48 (19), p.11453-11461
Main Authors: Green, Hyatt C, White, Karen M, Kelty, Cathy A, Shanks, Orin C
Format: Article
Language:English
Subjects:
Analysis methods
Animals
Applied sciences
Base Sequence
Biological and medical sciences
Biological Assay - methods
Biotechnology
Contamination
Cyclonic Storms
DNA, Bacterial - genetics
Dogs
Exact sciences and technology
Feces
Feces - microbiology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic Markers
Genetic technics
Genome
Genomics
Humans
Methods. Procedures. Technologies
Natural water pollution
Open Reading Frames - genetics
Pollution
Polymerase chain reaction
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Sewage - microbiology
Water Microbiology
Water Quality
Water treatment and pollution
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Summary: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 nonribosomal 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 noncanine hosts. As a proof-of-concept, these markers were detected in urban stormwater samples, suggesting a future application of newly developed methods for water quality monitoring.
ISSN:0013-936X
1520-5851
DOI:10.1021/es502637b