title: Detection of respiratory viruses by real-time polymerase chain reaction in outpatients with acute respiratory infection.
abstract: Viruses are the major contributors to the morbidity and mortality of upper and lower acute respiratory infections (ARIs) for all age groups. The aim of this study was to determine the frequencies for a large range of respiratory viruses using a sensitive molecular detection technique in specimens from outpatients of all ages with ARIs. Nasopharyngeal aspirates were obtained from 162 individuals between August 2007-August 2009. Twenty-three pathogenic respiratory agents, 18 respiratory viruses and five bacteria were investigated using multiplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) and indirect immunofluorescence assay (IIF). Through IIF, 33 (20.4%) specimens with respiratory virus were recognised, with influenza virus representing over half of the positive samples. Through a multiplex real-time RT-PCR assay, 88 (54.3%) positive samples were detected; the most prevalent respiratory viral pathogens were influenza, human rhinovirus and respiratory syncytial virus (RSV). Six cases of viral co-detection were observed, mainly involving RSV. The use of multiplex real-time RT-PCR increased the viral detection by 33.9% and revealed a larger number of respiratory viruses implicated in ARI cases, including the most recently described respiratory viruses [human bocavirus, human metapneumovirus, influenza A (H1N1) pdm09 virus, human coronavirus (HCoV) NL63 and HCoV HKU1].
authors.affiliation: Laboratório de Vírus Respiratórios, Núcleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, ES, Brasil.
authors.forename: Ronaldo Bragança
authors.initials: RB
authors.lastname: Martins Júnior
authors.affiliation: Laboratório de Vírus Respiratórios, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
authors.forename: Sharon
authors.initials: S
authors.lastname: Carney
authors.affiliation: Laboratório de Vírus Respiratórios, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Goldemberg
authors.affiliation: Laboratório Central de Saúde Pública do Espírito Santo, Vitória, ES, Brasil.
authors.forename: Lucas
authors.initials: L
authors.lastname: Bonine
authors.affiliation: Laboratório de Vírus Respiratórios, Núcleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, ES, Brasil.
authors.forename: Liliana Cruz
authors.initials: LC
authors.lastname: Spano
authors.affiliation: Laboratório de Vírus Respiratórios, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
authors.forename: Marilda
authors.initials: M
authors.lastname: Siqueira
authors.affiliation: Laboratório de Vírus Respiratórios, Núcleo de Doenças Infecciosas, Universidade Federal do Espírito Santo, Vitória, ES, Brasil.
authors.forename: Rita Elizabeth
authors.initials: RE
authors.lastname: Checon
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D000328:Adult; D017677:Age Distribution; D000368:Aged; D002648:Child; D002675:Child, Preschool; D017934:Coronavirus / Q000302:isolation & purification; D005260:Female; D056661:Human bocavirus / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009980:Influenza A virus / Q000302:isolation & purification*; D009981:Influenza B virus / Q000302:isolation & purification*; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009305:Nasopharynx / Q000821:virology; D010045:Outpatients; D010252:Paramyxoviridae / Q000302:isolation & purification*; D010849:Picornaviridae / Q000302:isolation & purification; D015995:Prevalence; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000821:virology*; D055815:Young Adult