Comparison of two poultry litter qPCR assays targeting the 16S rRNA gene of Brevibacterium sp.
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文摘
Chicken feces commonly contain 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 poultry-associated fecal pollution. We tested both assays against chicken litter (n聽=聽40), chicken fecal samples (n聽=聽186), non-chicken fecal sources (n聽=聽484), and environmental water samples (n聽=聽323). Most chicken litter samples (i.e., 鈮?8%) were positive for both assays with relatively high signal intensities, whereas only 23% and 12% of poultry fecal samples (n聽=聽186) were positive with the LA35 and the CL assays, respectively. Data using fecal samples from non-target animal species showed that the assays are highly host-associated (鈮?5%). Bayesian statistical models showed that the two assays are associated with relatively 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.
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