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Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources
Citation:
Sinigalliano, C., J. Ervin, L. Van De Werfhorst, B. Badgley, E. Balleste', J. Bartkowiak, A. Boehm, M. Byappanahalli, K. Goodwin, M. Gourmelon, J. Griffith, P. Holden, J. Jay, B. Layton, C. Lee, J. Lee, W. Meijer, R. Noble, M. Raith, H. Ryu, M. Sadowsky, A. Schriewer, D. Wang, D. Wanless, R. Whitman, S. Wuertz, AND J. Santodomingo. Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources. WATER RESEARCH. Elsevier Science Ltd, New York, NY, 47(18):6883–6896, (2013).
Impact/Purpose:
To inform the public of predictive tools for sustainable solutions for waterborne chemical and microbial contaminants.
Description:
Here we report results from a multi-laboratory (n=11) evaluation of four different PCR methods targeting the 16S rRNA gene of Catellicoccus marimammalium used to detect fecal contamination from birds in coastal environments. The methods included conventional end-point PCR, a SYBR® Green qPCR method, and two TaqMan® qPCR methods. Data processing had a pronounced impact on assay sensitivity and specificity calculations. Across laboratory standardization of metrics including the lower limit of quantification (LLOQ), target detected but not quantifiable (DNQ), and target not detected (ND) significantly improved results compared to results submitted by individual laboratories prior to definition standardization. The procedure used for data normalization also had a pronounced effect on perceived performance. The MST methods tested here were designed for gulls but also detected feces from other birds, particularly feces from pigeons found at the coast. Pigeons were found to contain sequences similar to C. marimammalium from gull feces. This study represents an important first step in the multilaboratory assessment of these methods and highlights the need to broaden and standardize additional evaluations, including environmentally relevant target concentrations in ambient waters from diverse geographic regions.
