Diagnostic discrimination of live attenuated influenza vaccine strains and community-acquired pathogenic strains in clinical samples

Nikki E. Freed, Christopher A. Myers, Kevin L. Russell, Elizabeth A. Walter, Marina Irvine, Robert G. Coon, David Metzgar

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)103-110
Number of pages8
JournalMolecular and Cellular Probes
Issue number2
StatePublished - Apr 2007
Externally publishedYes


  • FluMist
  • LAIV
  • PCR
  • RT-PCR
  • Vaccine shedding

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


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