title: A new real-time RT-qPCR assay for the detection, subtyping and quantification of human respiratory syncytial viruses positive- and negative-sense RNAs.
abstract: Human respiratory syncytial virus (RSV) is a major health problem and the main cause of hospitalization due to bronchiolitis. RSV is divided into two antigenic subgroups, RSV-A and -B that co-circulate worldwide. Rapid and sensitive detection is desirable for proper patient handling while assessment of viral load may help to evaluate disease severity and progression. Finally RSV subtyping is needed to determine the prevalence and pathogenicity of each RSV subgroup, as well as their sensitivity to treatment. In this study, we took into account the most recent circulating RSV variants and designed two quantitative TaqMan one-step RT-PCR assays to detect and quantify both RSV subgroups separately. Standard dilutions of transcripts of positive and negative polarities were included in the assay validation to assess potential differences in sensitivity on negative-sense genomes and positive-sense RNAs. In addition, RSV detection in respiratory specimens of different types and sampled in different populations was compared to commercially available RSV diagnostic tools. Altogether, the RSV-A and -B assays revealed sensitive and quantitative over a wide range of viral loads, with a slight improved sensitivity of the RSV-B assay on positive sense transcripts, and allowed accurate RSV subtyping. We thus provide a useful tool for both RSV diagnostics and research.
authors.affiliation: Faculty of Medicine of Geneva, Department of Microbiology and Molecular medicine, 1 rue Michel Servet, 1211 Geneva 4, Switzerland; Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Manel.Essaidi-Laziosi@hcuge.ch.
authors.forename: Manel
authors.initials: M
authors.lastname: Essaidi-Laziosi
authors.affiliation: Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Matthieu.Lyon@etu.unige.ch.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Lyon
authors.affiliation: Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Aline.Mamin@hcuge.ch.
authors.forename: Aline
authors.initials: A
authors.lastname: Mamin
authors.affiliation: Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Melanie.fernandesRocha@hcuge.ch.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Fernandes Rocha
authors.affiliation: Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Laurent.Kaiser@hcuge.ch.
authors.forename: Laurent
authors.initials: L
authors.lastname: Kaiser
authors.affiliation: Faculty of Medicine of Geneva, Department of Microbiology and Molecular medicine, 1 rue Michel Servet, 1211 Geneva 4, Switzerland; Geneva University Hospitals, Infectious Diseases Divisions, Rue Gabrielle-Perret-Gentil 4, 1211 Geneva 4, Switzerland. Electronic address: Caroline.Tapparel@hcuge.ch.
authors.forename: Caroline
authors.initials: C
authors.lastname: Tapparel
mesh_terms: D016679:Genome, Viral; D006801:Humans; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D012367:RNA, Viral
keywords: Antigenome; Genome; One step real-time PCR; Quantitative; Respiratory syncytial virus