title: Evaluation of methods using celite to concentrate norovirus, adenovirus and enterovirus from wastewater.
abstract: Enteroviruses, noroviruses and adenoviruses are among the most common viruses infecting humans worldwide. These viruses are shed in the feces of infected individuals and can accumulate in wastewater, making wastewater a source of a potentially diverse group of enteric viruses. In this study, two procedures were evaluated to concentrate noroviruses, adenoviruses and enteroviruses from primary effluent of wastewater. In the first procedure, indigenous enteroviruses, noroviruses and adenoviruses were concentrated using celite (diatomaceous earth) followed by centrifugation through a 30K MWCO filter and nucleic acid extraction. The second procedure used celite concentration followed by nucleic acid extraction only. Virus quantities were measured using qPCR. A second set of primary effluent samples were seeded with Coxsackievirus A7, Coxsackievirus B1, poliovirus 1 or enterovirus 70 before concentration and processed through both procedures for recovery evaluation of enterovirus species representatives. The pairing of the single step extraction procedure with the celite concentration process resulted in 47-98% recovery of examined viruses, while the celite concentration process plus additional centrifugal concentration before nucleic acid extraction showed reduced recovery (14-47%). The celite concentration process followed by a large volume nucleic acid extraction technique proved to be an effective procedure for recovering these important human pathogens from wastewater.
authors.affiliation: Biohazard Assessment Research Branch, Microbiological and Chemical Exposure Assessment Research Division, National Exposure Research Laboratory, Office of Research and Development, United States Environmental Protection Agency, 26 West Martin Luther King Drive, Cincinnati, OH 45268, USA. brinkman.nichole@epa.gov
authors.forename: Nichole E
authors.initials: NE
authors.lastname: Brinkman
authors.forename: Tyler D
authors.initials: TD
authors.lastname: Haffler
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Cashdollar
authors.forename: Eric R
authors.initials: ER
authors.lastname: Rhodes
mesh_terms: D000260:Adenoviruses, Human / Q000302:isolation & purification*; D002498:Centrifugation / Q000379:methods; D007692:Diatomaceous Earth / Q000737:chemistry*; D004770:Enterovirus / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods; D006801:Humans; D029322:Norovirus / Q000302:isolation & purification*; D009696:Nucleic Acids / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D014773:Virology / Q000379:methods*; D062065:Wastewater / Q000821:virology*
chemical_list: D007692:Diatomaceous Earth; D009696:Nucleic Acids; D062065:Wastewater
keywords: Adenovirus; Celite; Enterovirus; Norovirus; Wastewater