Luminex detection of fecal indicators in river samples, marine recreational water, and beach sand.
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.
Baums
Iliana B
IB
Cooperative Institute for Marine and Atmospheric Studies, Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL 33149, USA.
Goodwin
Kelly D
KD
Kiesling
Traci
T
Wanless
David
D
Diaz
Mara R
MR
Fell
Jack W
JW
D001419:Bacteria / Q000235:genetics*; D001483:Base Sequence; D016000:Cluster Analysis; D017931:DNA Primers; D015342:DNA Probes; D004784:Environmental Monitoring / Q000379:methods*; D005243:Feces / Q000382:microbiology*; D005456:Fluorescent Dyes; D008863:Microspheres; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D045483:Rivers / Q000382:microbiology*; D012623:Seawater / Q000382:microbiology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012822:Silicon Dioxide / Q000032:analysis*
D017931:DNA Primers; D015342:DNA Probes; D005456:Fluorescent Dyes; D012336:RNA, Ribosomal, 16S; D012822:Silicon Dioxide