title: Applicability of a real-time quantitative PCR assay for diagnosis of respiratory syncytial virus infection in immunocompromised adults.
abstract: Respiratory syncytial virus (RSV) accounts for the majority of respiratory virus infections, producing high mortality rates in immunocompromised patients with hematologic malignancies. The available methods for the rapid detection of RSV by antigen detection or PCR either lack sensitivity, require complex laboratory manipulation, or have not been evaluated in this patient population. To assess the applicability of a TaqMan-based real-time PCR technique for the detection of RSV A and B in immunocompromised adults, we developed a rapid, sensitive detection method that simultaneously detects RSV A and B and can be applied in routine diagnostics. The specificity of the assay was assessed using a panel of reference strains of other respiratory viruses and RSV. Electron microscopy-counted stocks of RSV A and B were used to develop a quantitative PCR format. Eleven copies of viral RNA could be detected for RSV A strain Long, and 14 copies could be detected for RSV B strain 9320, corresponding to 50% tissue culture infective doses of 0.86 and 0.34, respectively. The assay was evaluated on 411 combined nose and throat swabs derived from immunocompromised adults with or without signs of respiratory tract infection. The diagnostic efficacy of the TaqMan PCR determined on the clinical samples showed that this real-time PCR technique was substantially more sensitive than the combination of conventional viral culture and shell vial culture. None of the clinical specimens derived from patients without signs of respiratory illness were found to be positive for RSV by real-time TaqMan PCR.
authors.affiliation: Department of Virology, University Medical Center Utrecht, Utrecht, The Netherlands. l.vanelden@azu.nl
authors.forename: L J R
authors.initials: LJ
authors.lastname: van Elden
authors.forename: A M
authors.initials: AM
authors.lastname: van Loon
authors.forename: A
authors.initials: A
authors.lastname: van der Beek
authors.forename: K A W
authors.initials: KA
authors.lastname: Hendriksen
authors.forename: A I M
authors.initials: AI
authors.lastname: Hoepelman
authors.forename: M G J
authors.initials: MG
authors.lastname: van Kraaij
authors.forename: P
authors.initials: P
authors.lastname: Schipper
authors.forename: M
authors.initials: M
authors.lastname: Nijhuis
mesh_terms: D000328:Adult; D006801:Humans; D016867:Immunocompromised Host; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis*; D018113:Respiratory Syncytial Virus, Human / Q000302:isolation & purification*; D012680:Sensitivity and Specificity