title: Effectiveness of an RT-booster-PCR method for detection of noroviruses in stools collected after an outbreak of gastroenteritis.
abstract: Noroviruses exhibit a wide genomic and antigenic diversity, making laboratory diagnosis difficult. The abrupt onset of the illness does often not allow timely sample collection. Three different methods (a commercially available ELISA, a published RT-PCR and an RT-booster-PCR) were compared for detecting noroviruses in stools collected after the end of a gastroenteritis outbreak. Both ELISA and the RT-PCR detected noroviruses in 6 samples out of the 41 samples collected and tested. The RT-booster-PCR, however, was able to detect noroviruses in 23 (56%) of the samples with 20 of the samples confirmed by sequencing. Confirmation of PCR products was also undertaken by Southern hybridisation with controversial results (e.g. lack of confirmation on samples positive by sequencing). The results show that common molecular diagnostic methods may fail sometimes to detect the presence of noroviruses in a relevant proportion of samples. A sensitive detection method, such as the RT-booster-PCR described below may resolve the cases.
authors.affiliation: Istituto Superiore di Sanità, Centro Nazionale per la Qualità degli Alimenti e per i Rischi Alimentari, Reparto Pericoli Microbiologici connessi con gli Alimenti, Viale Regina Elena 299, 00161 Roma, Italy. dario.demedici@iss.it
authors.forename: Dario
authors.initials: D
authors.lastname: De Medici
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Suffredini
authors.forename: Silvia
authors.initials: S
authors.lastname: Crudeli
authors.forename: Franco Maria
authors.initials: FM
authors.lastname: Ruggeri
mesh_terms: D017250:Caliciviridae Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D017931:DNA Primers; D004196:Disease Outbreaks; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D006801:Humans; D029322:Norovirus / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012367:RNA, Viral