title: Evaluation of the TRCRtest NV-W for norovirus detection in stools by the Transcription-Reverse Transcription Concerted method.
abstract: A novel molecular assay, TRCRtest NV-W, based on a transcription-reverse transcription concerted reaction (TRC) for isothermal amplification and real-time detection of norovirus in stools was assessed and compared with an RT-nPCR. Archived stools positive for either different types or variants of norovirus genogroups I and II or other enteric viruses were used to assess the sensitivity and specificity of the novel assay. The TRC assay was 100% specific since it detected all the noroviruses tested and it did not display cross reactivity with other enteric viruses. When screening a collection of 387 stools with the TRC and RT-nPCR assays, the TRC displayed concordance, sensitivity, specificity, positive and negative predictive values of 96.6%, 81%, 99.7%, 98.1%, and 96.3%, respectively, after retesting the negative specimens. Additional PCRs and/or sequencing, used to understand inconsistent results between TRC and RT-nPCR, confirmed all positive results and did not reveal nucleotide variations in the TRC probe and primers binding sites. The TRC assay may be a rapid and ease of use tool for the detection of noroviruses in clinical virology laboratories even in the face of rapidly evolving noroviruses.
authors.affiliation: Unit of Microbiology and Virology, Department of Clinical and Experimental Medicine, University of Parma, Viale Antonio Gramsci, 14, 43126, Parma, Italy. Electronic address: mariacristina.medici@unipr.it.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Medici
authors.forename: Fabio
authors.initials: F
authors.lastname: Tummolo
authors.forename: Valeria
authors.initials: V
authors.lastname: Albonetti
authors.forename: Federica
authors.initials: F
authors.lastname: Pinardi
authors.forename: Francesca
authors.initials: F
authors.lastname: Ferraglia
authors.forename: Carlo
authors.initials: C
authors.lastname: Chezzi
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Arcangeletti
authors.forename: Flora
authors.initials: F
authors.lastname: De Conto
authors.forename: Adriana
authors.initials: A
authors.lastname: Calderaro
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017250:Caliciviridae Infections / Q000175:diagnosis* / Q000821:virology; D002648:Child; D002675:Child, Preschool; D005243:Feces / Q000821:virology*; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic; D014773:Virology / Q000379:methods; D055815:Young Adult
chemical_list: D012367:RNA, Viral
keywords: EM; Enteritis; G; NoV; Norovirus; Rapid diagnosis; TRC; TRC method; electron microscopy; genogroup; norovirus; transcription-reverse transcription concerted reaction; v.; variant