title: Diagnostic discrimination of live attenuated influenza vaccine strains and community-acquired pathogenic strains in clinical samples.
abstract: Live vaccines can generate false-positive results on common influenza assays including reverse transcriptase-PCR (RT-PCR), culture and antigen tests. This threatens the integrity of epidemiological data and may misdirect treatment and control efforts. We report the development of RT-PCR tests that distinguish live FluMist vaccine (FMV) strains from circulating influenza strains in clinical samples. Primers were validated using influenza-positive samples from unvaccinated patients, packaged FMV, and one PCR-positive asymptomatic vaccine. Furthermore, the assay was used to experimentally test our lab's collection of influenza-positive samples from the 2004-05 and 2005-06 influenza seasons and several 2005 preseason isolates to determine the rate of vaccine-derived false-positive results under differing epidemiological conditions. Analytical and clinical validations show that the assay is both sensitive and specific. Experimental results demonstrate that 51 out of 51 influenza-positive samples collected during influenza season from ill, previously-vaccinated military personnel represent real infections with circulating strains. Finally, the assay shows that four preseason influenza-positive samples were false positives resulting from vaccine shedding. The vaccine-discriminatory RT-PCR methods described here provide the first test designed to distinguish FMV strains from circulating strains. The results show that the test is effective, and demonstrate the importance of such tests in the age of live vaccines.
authors.affiliation: Department of Defense Center for Deployment Health Research, Naval Health Research Center, San Diego, CA 92186-5122, USA. nicole.freed@env.ethz.ch
authors.forename: Nikki E
authors.initials: NE
authors.lastname: Freed
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Myers
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Russell
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Walter
authors.forename: Marina
authors.initials: M
authors.lastname: Irvine
authors.forename: Robert G
authors.initials: RG
authors.lastname: Coon
authors.forename: David
authors.initials: D
authors.lastname: Metzgar
mesh_terms: D001483:Base Sequence; D017714:Community-Acquired Infections / Q000276:immunology*; D017931:DNA Primers; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000472:pathogenicity*; D007252:Influenza Vaccines / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014613:Vaccines, Attenuated
chemical_list: D017931:DNA Primers; D007252:Influenza Vaccines; D014613:Vaccines, Attenuated