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Appl. Environ. Microbiol. doi:10.1128/AEM.02546-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Simple and Rapid F+ Coliphage Culture, Latex Agglutination, and Typing (CLAT) Assay to Detect and Source Track Fecal Contamination

Department of Environmental Sciences and Engineering, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599

^* To whom correspondence should be addressed. Email: dlove{at}email.unc.edu.

	Abstract

Simple, rapid and reliable fecal indicator tests are needed to better monitor and manage ambient waters and treated waters and wastes. Antibody-coated polymeric bead agglutination assays can fulfill these needs, and are inexpensive, field-portable for non-lab settings, and their reagents can be stored at ambient temperatures for months. The goal of this study was to develop, optimize, and validate a rapid microbial water quality monitoring assay using F+ coliphage culture, latex agglutination and typing (CLAT) to detect F+ coliphage groups with antibody-coated particles. Rapid (180 minute) F+ coliphage culture was comparable to 16-24 hour culture time used in EPA Method 1601 and was amenable to CLAT detection. CLAT was performed on a cardboard card by mixing a drop of coliphage enrichment culture with a drop of antibody-coated polymeric beads as the detection reagent. Visual agglutination or clumping of positive samples occurred in <60 seconds. The CLAT assay had a sensitivity of 96.4% (185/192 samples) and 98.2% (161/164 samples), and a specificity of 100% (34/34 samples) and 97.7% (129/132 samples) for F+ RNA and DNA coliphages, respectively. CLAT successfully identified F+ RNA coliphages into serogroups typically from human (groups II/III) and animal (groups I/IV) fecal sources, and in similar proportions as a nucleic acid hybridization assay. This novel group-specific antibody-based particle agglutination technique for rapid and simple detection and grouping of F+ coliphages provides a new and improved tool to monitor the microbiological quality of drinking, recreational, shellfishing, and other waters.