Simple and rapid F+ coliphage culture, latex agglutination, and typing assay to detect and source track fecal contamination.
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 and portable for nonlaboratory 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 min) F+ coliphage culture gave comparable results to those with the 16- to 24-h culture time used in EPA method 1601 and was amenable to CLAT assay 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 sensitivities of 96.4% (185/192 samples) and 98.2% (161/164 samples) and specificities of 100% (34/34 samples) and 97.7% (129/132 samples) for F+ RNA and DNA coliphages, respectively. CLAT successfully classified F+ RNA coliphages into serogroups typically obtained from human (groups II and III) and animal (groups I and IV) fecal sources, in similar proportions to those obtained with 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 for monitoring the microbiological quality of drinking, recreational, shellfishing, and other waters.
Love
David C
DC
Department of Environmental Sciences and Engineering, School of Public Health, University of North Carolina, Chapel Hill, NC 27599, USA. dlove@email.unc.edu
Sobsey
Mark D
MD
D001434:Bacteriophage Typing / Q000379:methods*; D001483:Base Sequence; D003090:Coliphages / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005144:F Factor / Q000235:genetics; D005243:Feces / Q000821:virology*; D006801:Humans; D066298:In Vitro Techniques; D007841:Latex Fixation Tests / Q000379:methods*; D017909:Levivirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D014776:Virus Cultivation / Q000379:methods*; D014871:Water Microbiology
D017931:DNA Primers; D004279:DNA, Viral