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Comparison of two poultry litter qPCR assays targeting the 16S rRNA gene of Brevibacterium sp
Ryu, H., M. Elk, I. Khan, V. Harwood, M. Molina, T. Edge, AND J. Santodomingo. Comparison of two poultry litter qPCR assays targeting the 16S rRNA gene of Brevibacterium sp. WATER RESEARCH. Elsevier Science Ltd, New York, NY, 48(1):613-621, (2014).
Chicken feces are vectors of human pathogens and are also important sources of fecal pollution in environmental waters. Consequently, methods that can detect chicken fecal pollution are needed in public health and environmental monitoring studies. In this study, we compared a previously developed SYBR green qPCR assay (LA35) to a novel TaqMan qPCR assay (CL) for the environmental detection of chicken-specific fecal pollution. We tested both assays against chicken litter (n=40) and chicken fecal samples (n=186), non-chicken fecal sources (n=484), and environmental water samples (n=323). Most chicken litter samples (i.e., ≥98%) were positive for both assays with relatively high signal intensities, whereas only 23% and 12% of poultry fecal samples (n=186) were positive with the LA35 and the CL assays, respectively. Data using fecal samples from non-targeted animal species showed that the assays are highly host-specific (≥95%). Bayesian statistical models showed that two assays are associated with significantly low probability of false-positive and false-negative signals in water samples. The CL marker had a lower prevalence than the LA35 assay when tested against environmental water samples (i.e., 21% vs. 31% positive signals). However, by combining the results from the two assays the detection levels increased to 41%, suggesting that using multiple assays can improve the detection of chicken-fecal pollution in environmental waters.
This study shows the importance of using multiple chicken poultry assays in environmental applications
Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL RISK MANAGEMENT RESEARCH LABORATORY
WATER SUPPLY AND WATER RESOURCES DIVISION
MICROBIAL CONTAMINANTS CONTROL BRANCH