Luminex detection of fecal indicators in river samples, marine recreational water, and beach sand.pdfVIP

/locate/marpolbul Marine Pollution Bulletin 54 (2007) 521–536Luminex detection of fecal indicators in river samples, marine recreational water, and beach sand Iliana B. Baums a,1, Kelly D. Goodwin b,*, Traci Kiesling a, David Wanless a, Mara R. Diaz c, Jack W. Fell c a Cooperative Institute for Marine and Atmospheric Studies, Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL 33149, USA b National Oceanographic and Atmospheric Administration, Atlantic Oceanographic and Meteorological Laboratory, Ocean Chemistry Division, Miami, FL 33149, USA c Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL 33149, USAAbstract Research to understand and remediate coastal pollution is moving toward a multitiered approach in which traditional enumeration of fecal indicators is accompanied by molecular analysis of a variety of targets. Technology that rapidly detects multiple microbial contam- inants would benefit from such an approach. The Luminex 100TM system is a suspension array that assays multiple analytes rapidly in a single well of a microtiter plate. The ability of the system to simultaneously detect multiple fecal indicating bacteria in environmental samples was tested. Primer/probe sets were designed to simultaneously detect the following fecal indicators: the Bacteroides fragilis group, Enterococcus spp., Escherichia coli and Shigella spp., Bacteroides distasonis, and Ent. faecalis. Specificity and sensitivity of the Luminex probes was tested against laboratory cultures. In addition, sequencing, culture plate testing, and specificity testing with envi- ronmental isolates were steps taken to validate the function of the assay with environmental samples. Luminex response to cultures and to environmental samples was consistent with sequencing results, suggesting that the technology has the potential to simultaneously detect multiple targets for coastal water quality applications,