title: Rapid virus detection procedure for molecular tracing of shellfish associated with disease outbreaks.
abstract: Detection of pathogenic viruses in oysters implicated in gastroenteritis outbreaks is often hampered by time-consuming, specialist virus extraction methods. Five virus RNA extraction methods were evaluated with respect to performance characteristics and sensitivity on artificially contaminated oyster digestive glands. The two most promising procedures were further evaluated on bioaccumulated and naturally contaminated oysters. The most efficient method was used to trace the source in an outbreak situation. Out of five RNA extraction protocols, PEG precipitation and the RNeasy Kit performed best with norovirus genogroup III-spiked digestive glands. Analyzing 24-h bioaccumulated oysters revealed a slightly better sensitivity with PEG precipitation, but the RNeasy Kit was less prone to concentrate inhibitors. The latter procedure demonstrated the presence of human noroviruses in naturally contaminated oysters and oysters implicated in an outbreak. In this outbreak, in four out of nine individually analyzed digestive glands, norovirus was detected. In one of the oysters and in one of the fecal samples of the clinical cases, identical norovirus strains were detected. A standard and rapid virus extraction method using the RNeasy Kit appeared to be most useful in tracing shellfish as the source in gastroenteritis outbreaks, and to be able to make effective and timely risk management decisions.
authors.affiliation: Microbiological Laboratory for Health Protection, National Institute of Public Health and the Environment, P.O. Box 1, NL-3720 BA Bilthoven, The Netherlands.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: de Roda Husman
authors.forename: Froukje
authors.initials: F
authors.lastname: Lodder-Verschoor
authors.forename: Harold H J L
authors.initials: HH
authors.lastname: van den Berg
authors.forename: Françoise S
authors.initials: FS
authors.lastname: Le Guyader
authors.forename: Hilde
authors.initials: H
authors.lastname: van Pelt
authors.forename: Wim H M
authors.initials: WH
authors.lastname: van der Poel
authors.forename: Saskia A
authors.initials: SA
authors.lastname: Rutjes
mesh_terms: D000818:Animals; D011232:Chemical Precipitation; D004196:Disease Outbreaks; D005243:Feces / Q000821:virology; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D010125:Ostreidae / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis; D011933:Reagent Kits, Diagnostic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000821:virology*; D014780:Viruses / Q000302:isolation & purification* / Q000472:pathogenicity
chemical_list: D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic