Interlaboratory comparison of real-time pcr protocols for quantification of general fecal indicator bacteria

摘要: The application of quantitative real-time PCR (qPCR) technologies for the rapid identification fecal bacteria in environmental waters is being considered use as a national water quality metric United States. transition from research tool to standardized protocol requires information on reproducibility and sources variation associated with qPCR methodology across laboratories. This study examines interlaboratory variability measurement enterococci Bacteroidales concentrations standardized, spiked, DNA using Entero1a GenBac3 methods, respectively. Comparisons are based data generated eight different facilities. Special attention was placed influence isolation step effect simplex multiplex amplification approaches variability. Results suggest that crude lysate sufficient unless samples contain substances can inhibit amplification. No appreciable difference observed between approaches. Overall, levels remained low (<10% coefficient variation) regardless protocol.