title: Monitoring epidemic viral respiratory infections using one-step real-time triplex RT-PCR targeting influenza A and B viruses and respiratory syncytial virus.
abstract: Rapid and specific diagnosis of influenza A/B and respiratory syncytial virus (RSV) viruses is needed for optimal management of patients with acute respiratory infections. In this study, a one-step triplex real-time RT-PCR assay was developed for rapid diagnosis of influenza A/B and RSV infections to optimize diagnosis efficiency of acute respiratory infections. Cell-culture supernatants and clinical samples were used to evaluate specificity and sensitivity of the assay. The assay was used routinely during two winter epidemics for testing respiratory specimens from 2,417 patients. The limit of detection in cell-culture supernatant was 1-10 plaque forming units/input (influenza A/B) and 2 × 10(-2) 50% tissue culture infectious dose/input (RSV). In clinical samples, the assay was as sensitive as commercial molecular assays for the detection of each influenza A/B and RSV (Flu-A/B and RSV-A/B r-gene™) individually, and far more sensitive than antigen detection. During the winter 2008-2009, the assay identified 145 RSV, 42 influenza A, and one mixed RSV-influenza A infections among 298 patients. The next winter, the assay was used in two independent hospital laboratory settings. 776 patients were tested in one hospital and 1,343 in the other, resulting in 184 and 501 RSV, 133 and 150 influenza A, and 1 and 11 mixed RSV-influenza A infections, respectively, being detected. This new user-friendly assay allows rapid (within hours), effective molecular diagnosis of single or mixed infections involving influenza A (including seasonal A H1N1 and H3N2, and A(H1N1) 2009), influenza B, and RSV(A/B). The assay is very valuable for managing patients during winter epidemics when influenza and respiratory syncytial viruses co-circulate.
authors.affiliation: AP-HP, Ambroise Paré Hospital, Department of Pediatrics, Boulogne-Billancourt, France.
authors.forename: Solesne
authors.initials: S
authors.lastname: Papillard-Marechal
authors.forename: Vincent
authors.initials: V
authors.lastname: Enouf
authors.forename: Aurélie
authors.initials: A
authors.lastname: Schnuriger
authors.forename: Astrid
authors.initials: A
authors.lastname: Vabret
authors.forename: Edouard
authors.initials: E
authors.lastname: Macheras
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Rameix-Welti
authors.forename: Bernard
authors.initials: B
authors.lastname: Page
authors.forename: François
authors.initials: F
authors.lastname: Freymuth
authors.forename: Sylvie
authors.initials: S
authors.lastname: van der Werf
authors.forename: Antoine
authors.initials: A
authors.lastname: Garbarg-Chenon
authors.forename: Bertrand
authors.initials: B
authors.lastname: Chevallier
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Gaillard
authors.forename: Elyanne
authors.initials: E
authors.lastname: Gault
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000302:isolation & purification*; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology