title: Molecular detection of human adenovirus in sediment using a direct detection method compared to the classical polyethylene glycol precipitation.
abstract: Various effective methods have been developed to measure the concentration of viruses in sediment samples. However, there is need to standardize less laborious and simpler techniques. The objective of the present study was to compare two different methods to measure the concentration of viruses in soil samples. The use of polyethylene glycol (PEG) was compared with a direct extraction of viral nucleic acids from the samples diluted in modified Eagle's minimal essential medium (E-MEM). The presence of adenovirus in the samples was detected by real-time quantitative polymerase chain reaction (qPCR). Only six samples (30%) were positive for adenovirus when PEG technique was used. The direct method showed 16 (80%) samples positive for adenovirus. Therefore, direct detection (i.e. without previous concentration) demonstrated a higher rate of detection, better effectiveness, and shorter execution time. Furthermore, direct detection uses reagents that are often readily available in virology laboratories. Thus, it is an attractive alternative to other methods of detection of virus particles in sediments.
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil. Electronic address: rstaggemeier@gmail.com.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Staggemeier
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil.
authors.forename: Marina
authors.initials: M
authors.lastname: Bortoluzzi
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil.
authors.forename: Tatiana Moraes
authors.initials: TM
authors.lastname: da Silva Heck
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil.
authors.forename: Tiago
authors.initials: T
authors.lastname: da Silva
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil.
authors.forename: Fernando Rosado
authors.initials: FR
authors.lastname: Spilki
authors.affiliation: Laboratory of Molecular Microbiology, Universidade Feevale, ERS 239, No. 2755, Novo Hamburgo, Rio Grande do Sul 93352-000, Brazil.
authors.forename: Sabrina Esteves
authors.initials: SE
authors.lastname: de Matos Almeida
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification*; D019015:Geologic Sediments / Q000821:virology*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012987:Soil; D014773:Virology / Q000379:methods*
chemical_list: D004279:DNA, Viral; D012987:Soil
keywords: Direct method; Enteric viruses; Sediment