title: Field-based evaluation of a male-specific (F+) RNA coliphage concentration method.
abstract: Fecal contamination of water poses a significant risk to public health due to the potential presence of pathogens, including enteric viruses. Therefore, sensitive, reliable and easy to use methods for the concentration, detection and quantification of microorganisms associated with the safety and quality of water are needed. In this study, we performed a field evaluation of an anion exchange resin-based method to concentrate male-specific (F+) RNA coliphages (FRNA), fecal indicator organisms, from diverse environmental waters that were suspected to be contaminated with feces. In this system, FRNA coliphages are adsorbed to anion exchange resin and direct nucleic acid isolation is performed, yielding a sample amenable to real-time reverse transcriptase (RT)-PCR detection. Matrix-dependent inhibition of this method was evaluated using known quantities of spiked FRNA coliphages belonging to four genogroups (GI, GII, GII and GIV). RT-PCR-based detection was successful in 97%, 72%, 85% and 98% of the samples spiked (106pfu/l) with GI, GII, GIII and GIV, respectively. Differential FRNA coliphage genogroup detection was linked to inhibitors that altered RT-PCR assay efficiency. No association between inhibition and the physicochemical properties of the water samples was apparent. Additionally, the anion exchange resin method facilitated detection of naturally present FRNA coliphages in 40 of 65 environmental water samples (61.5%), demonstrating the viability of this system to concentrate FRNA coliphages from water.
authors: National Wildlife Research Center, Wildlife Services, Animal and Plant Health Inspection Service, United States Department of Agriculture, Fort Collins, CO, USA.
authors: J C
authors: JC
authors: Chandler
authors: Department of Animal Sciences, Colorado State University, Fort Collins, CO, USA.
authors: A
authors: A
authors: Pérez-Méndez
authors: United States Environmental Protection Agency, New England Regional Laboratory, North Chelmsford, MA, USA.
authors: J
authors: J
authors: Paar
authors: ESAT Contractor-TechLaw, Inc., United States Environmental Protection Agency, New England Regional Laboratory, North Chelmsford, MA, USA.
authors: M M
authors: MM
authors: Doolittle
authors: Department of Animal Science, University of Wyoming, Laramie, WY, USA.
authors: B
authors: B
authors: Bisha
authors: Department of Food Science and Agricultural Chemistry, McGill University, Ste. Anne de Bellevue, Quebec, Canada. Electronic address: lawrence.goodridge@mcgill.ca.
authors: L D
authors: LD
authors: Goodridge
mesh_terms: D000327:Adsorption; D000837:Anion Exchange Resins / Q000191:economics; D003090:Coliphages / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D004784:Environmental Monitoring / Q000379:methods; D005144:F Factor; D005243:Feces / Q000821:virology; D006801:Humans; D017907:Leviviridae / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014871:Water Microbiology; D014876:Water Pollution / Q000032:analysis
chemical_list: D000837:Anion Exchange Resins; D012367:RNA, Viral