title: Evaluation of an anion exchange resin-based method for concentration of F-RNA coliphages (enteric virus indicators) from water samples.
abstract: Enteric viral contaminants in water represent a public health concern, thus methods for detecting these viruses or their indicator microorganisms are needed. Because enteric viruses and their viral indicators are often found at low concentrations in water, their detection requires upfront concentration methods. In this study, a strong basic anion exchange resin was evaluated as an adsorbent material for the concentration of F-RNA coliphages (MS2, Qβ, GA, and HB-P22). These coliphages are recognized as enteric virus surrogates and fecal indicator organisms. Following adsorption of the coliphages from 50ml water samples, direct RNA isolation and real time RT-PCR detection were performed. In water samples containing 10(5)pfu/ml of the F-RNA coliphages, the anion exchange resin (IRA-900) adsorbed over 96.7% of the coliphages present, improving real time RT-PCR detection by 5-7 cycles compared to direct testing. F-RNA coliphage RNA recovery using the integrated method ranged from 12.6% to 77.1%. Resin-based concentration of samples with low levels of the F-RNA coliphages allowed for 10(0)pfu/ml (MS2 and Qβ) and 10(-1)pfu/ml (GA and HB-P22) to be detected. The resin-based method offers considerable advantages in cost, speed, simplicity and field adaptability.
authors.affiliation: Department of Animal Sciences, Colorado State University, Fort Collins, CO, United States.
authors.forename: A
authors.initials: A
authors.lastname: Pérez-Méndez
authors.affiliation: Department of Animal Science, University of Wyoming, Laramie, WY, United States.
authors.forename: J C
authors.initials: JC
authors.lastname: Chandler
authors.affiliation: Department of Animal Science, University of Wyoming, Laramie, WY, United States.
authors.forename: B
authors.initials: B
authors.lastname: Bisha
authors.affiliation: Department of Food Science and Agricultural Chemistry, McGill University, Ste. Anne de Bellevue, Quebec, Canada. Electronic address: lawrence.goodridge@mcgill.ca.
authors.forename: L D
authors.initials: LD
authors.lastname: Goodridge
mesh_terms: D000818:Animals; D000837:Anion Exchange Resins / Q000191:economics; D003090:Coliphages / Q000302:isolation & purification*; D003365:Costs and Cost Analysis; D006801:Humans; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods; D013997:Time Factors; D014871:Water Microbiology; D014876:Water Pollution
chemical_list: D000837:Anion Exchange Resins; D012367:RNA, Viral
keywords: Anion exchange resin; Enteric virus indicators; F-RNA coliphages; Real time RT-PCR; Virus concentration methods