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

• Published in: Marine pollution bulletin Vol. 54; no. 5; pp. 521 - 536
• Main Authors: Baums, Iliana B., Goodwin, Kelly D., Kiesling, Traci, Wanless, David, Diaz, Mara R., Fell, Jack W.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.05.2007; Elsevier
• Subjects: Analysis methods; Applied sciences; Bacteria - genetics; Bacteroides; Bacteroides distasonis; Bacteroides fragilis; Base Sequence; Beaches; Brackish; Cluster Analysis; DNA Primers; DNA Probes; Earth sciences; Earth, ocean, space; Engineering and environment geology. Geothermics; Enterococcus; Environmental Monitoring - methods; Escherichia coli; Exact sciences and technology; Fecal indicating bacteria; Feces - microbiology; Fluorescent Dyes; Freshwater; Luminex; Marine; Microspheres; Molecular Sequence Data; Natural water pollution; Phylogeny; Pollution; Pollution, environment geology; Polymerase Chain Reaction; Recreational water quality; Rivers - microbiology; RNA, Ribosomal, 16S - genetics; Seawater - microbiology; Sensitivity and Specificity; Sequence Analysis, DNA; Silicon Dioxide - analysis; Water treatment and pollution; Fecal indicating bacteria; Recreational water quality; Beaches; Luminex; Bacteroides; Multiplexing; Sand; Microbiology; Escherichia coli; Coastal zone; Bacteroidaceae; Immunological method; Pathogenic; Micrococcales; Bacteria; Stream; Feces; Biological contamination; Enterobacteriaceae; Seawater; Enterococcus; Littoral zone; Biological indicator; Streptococcaceae; Analysis method; Surveillance; Fluorescent labelling; Surface water; Water quality; Water pollution; Immunofluorescence; Bathing water; Shigella
• ISSN: 0025-326X; 1879-3363
• DOI: 10.1016/j.marpolbul.2006.12.018

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 contaminants would benefit from such an approach. The Luminex ® 100™ 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 environmental 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, particularly as progress is made to efficiently extract DNA from water and sediment matrices.