title: Evaluation of a broadly reactive nucleic acid sequence based amplification assay for the detection of noroviruses in faecal material.
abstract: A recently described nucleic acid sequence based amplification (NASBA) assay for the detection of genogroup I (GI) and genogroup II (GII) norovirus RNA in faecal samples was evaluated against a reverse transcription polymerase chain reaction (RT-PCR). Both assays were used to screen a panel of 38 faecal samples known to contain 17 different norovirus strains and 131 clinical samples collected from 60 gastroenteritis outbreaks of unknown aetiology. The NASBA assay detected 13 out of the 17 strains of norovirus in the characterised panel, failing to detect a single GII strain and three GI strains. There was 90% agreement between the two assays used to detect norovirus in clinical samples from outbreaks. NASBA detected norovirus RNA in all 64 samples positive by RT-PCR and also detected norovirus RNA in additional 13 samples that were negative by RT-PCR. The sensitivity and specificity of NASBA was 100% and 80%, respectively, compared to RT-PCR results. The norovirus NASBA assay was shown to be highly sensitive and specific, and its ease of use and rapid turnaround time makes it a favourable alternative to RT-PCR for the investigation of norovirus outbreaks.
authors.affiliation: National Public Health Service for Wales Microbiology Cardiff, Specialist Virology Centre, University Hospital of Wales, Heath Park, Cardiff CF14 4XW, UK. catherine.moore@nphs.wales.nhs.uk
authors.forename: C
authors.initials: C
authors.lastname: Moore
authors.forename: E M
authors.initials: EM
authors.lastname: Clark
authors.forename: C I
authors.initials: CI
authors.lastname: Gallimore
authors.forename: S A
authors.initials: SA
authors.lastname: Corden
authors.forename: J J
authors.initials: JJ
authors.lastname: Gray
authors.forename: D
authors.initials: D
authors.lastname: Westmoreland
mesh_terms: D017250:Caliciviridae Infections / Q000821:virology*; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000821:virology*; D006801:Humans; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021161:Self-Sustained Sequence Replication; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral