title: From Lab to Lake - Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area.
abstract: Quantitative PCR methods are commonly used to monitor enteric viruses in the aquatic environment because of their high sensitivity, short reaction times and relatively low operational cost. However, conclusions for public health drawn from results of such molecular techniques are limited due to their inability to determine viral infectivity. Ethidium monoazide (EMA) and propidium monoazide (PMA) are capable to penetrate the damaged or compromised capsid of the inactivated viruses and bind to the viral nucleic acids. We assessed whether dye treatment is a suitable approach to improve the ability of qPCR to distinguish between infectious and non-infectious human adenovirus, enterovirus and rotavirus A in surface water of an urban river and sewage before and after UV disinfection. Like the gold standard of cell culture assays, pretreatment EMA-/PMA-qPCR succeeded in removing false positive results which would lead to an overestimation of the viral load if only qPCR of the environmental samples was considered. A dye pretreatment could therefore provide a rapid and relatively inexpensive tool to improve the efficacy of molecular quantification methods in regards to viral infectivity.
authors.affiliation: Ruhr-University Bochum, Department of Hygiene, Social- and Environmental Medicine, Bochum, Germany.
authors.forename: Mats
authors.identifier: http://orcid.org/0000-0002-1295-3410
authors.initials: M
authors.lastname: Leifels
authors.affiliation: Environmental Virology Laboratory, Department of Water Pollution Research, National Research Centre, Cairo, Egypt.
authors.forename: Ibrahim Ahmed
authors.initials: IA
authors.lastname: Hamza
authors.affiliation: Ruhr-University Bochum, Department of Hygiene, Social- and Environmental Medicine, Bochum, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Krieger
authors.affiliation: Ruhr-University Bochum, Department of Hygiene, Social- and Environmental Medicine, Bochum, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Wilhelm
authors.affiliation: University of Duisburg-Essen, Faculty of Chemistry, Biofilm Centre - Aquatic Microbiology, Essen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Mackowiak
authors.affiliation: Ruhr-University Bochum, Department of Hygiene, Social- and Environmental Medicine, Bochum, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Jurzik
mesh_terms: D001386:Azides / Q000737:chemistry; D004770:Enterovirus / Q000235:genetics*; D006801:Humans; D060106:Lakes; D011419:Propidium / Q000031:analogs & derivatives / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D045483:Rivers / Q000821:virology*; D012722:Sewage / Q000821:virology*; D014506:Urban Renewal; D014871:Water Microbiology
chemical_list: D001386:Azides; D012722:Sewage; C533957:propidium monoazide; D011419:Propidium; C014535:8-azidoethidium