1887

Abstract

False-positive PCR results usually occur as a consequence of specimen-to-specimen or amplicon-to-specimen contamination within the laboratory. Evidence of contamination at time of specimen collection linked to influenza vaccine administration in the same location as influenza sampling is described. Clinical, circumstantial and laboratory evidence was gathered for each of five cases of influenza-like illness (ILI) with unusual patterns of PCR reactivity for seasonal H1N1, H3N2, H1N1 (2009) and influenza B viruses. Two 2010 trivalent influenza vaccines and environmental swabs of a hospital influenza vaccination room were also tested for influenza RNA. Sequencing of influenza A matrix (M) gene amplicons from the five cases and vaccines was undertaken. Four 2009 general practitioner (GP) specimens were seasonal H1N1, H3N2 and influenza B PCR positive. One 2010 GP specimen was H1N1 (2009), H3N2 and influenza B positive. PCR of 2010 trivalent vaccines showed high loads of detectable influenza A and B RNA. Sequencing of the five specimens and vaccines showed greatest homology with the M gene sequence of Influenza A/Puerto Rico/8/1934 H1N1 virus (used in generation of influenza vaccine strains). Environmental swabs had detectable influenza A and B RNA. RNA detection studies demonstrated vaccine RNA still detectable for at least 66 days. Administration of influenza vaccines and clinical sampling in the same room resulted in the contamination with vaccine strains of surveillance swabs collected from patients with ILI. Vaccine contamination should therefore be considered, particularly where multiple influenza virus RNA PCR positive signals (e.g. H1N1, H3N2 and influenza B) are detected in the same specimen.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.036178-0
2012-03-01
2020-01-23
Loading full text...

Full text loading...

/deliver/fulltext/jmm/61/3/332.html?itemId=/content/journal/jmm/10.1099/jmm.0.036178-0&mimeType=html&fmt=ahah

References

  1. Aslanzadeh J. . ( 2004; ). Preventing PCR amplification carryover contamination in a clinical laboratory. . Ann Clin Lab Sci 34:, 389–396.[PubMed]
    [Google Scholar]
  2. Borst A. , Box A. T. , Fluit A. C. . ( 2004; ). False-positive results and contamination in nucleic acid amplification assays: suggestions for a prevent and destroy strategy. . Eur J Clin Microbiol Infect Dis 23:, 289–299. [CrossRef] [PubMed]
    [Google Scholar]
  3. Diallo I. S. , Read A. J. , Kirkland P. D. . ( 2011; ). Potential of vaccination to confound interpretation of real-time PCR results for equine influenza. . Vet Rec 169:, 252. [CrossRef] [PubMed]
    [Google Scholar]
  4. Ellis J. S. , Curran M. D. . ( 2011; ). Simultaneous molecular detection and confirmation of influenza AH5, with internal control. . Methods Mol Biol 665:, 161–181. [CrossRef] [PubMed]
    [Google Scholar]
  5. Freed N. E. , Myers C. A. , Russell K. L. , Walter E. A. , Irvine M. , Coon R. G. , Metzgar D. . ( 2007; ). Diagnostic discrimination of live attenuated influenza vaccine strains and community-acquired pathogenic strains in clinical samples. . Mol Cell Probes 21:, 103–110. [CrossRef] [PubMed]
    [Google Scholar]
  6. Geeraedts F. , Goutagny N. , Hornung V. , Severa M. , de Haan A. , Pool J. , Wilschut J. , Fitzgerald K. A. , Huckriede A. . ( 2008; ). Superior immunogenicity of inactivated whole virus H5N1 influenza vaccine is primarily controlled by toll like receptor signalling. . PLoS Pathog 4:, e1000138. [CrossRef]
    [Google Scholar]
  7. Greatorex J. S. , Page R. F. , Curran M. D. , Digard P. , Enstone J. E. , Wreghitt T. , Powell P. P. , Sexton D. W. , Vivancos R. , Nguyen-Van-Tam J. S. . ( 2010; ). Effectiveness of common household cleaning agents in reducing the viability of human influenza A/H1N1. . PLoS ONE 5:, e8987. [CrossRef] [PubMed]
    [Google Scholar]
  8. Hartley J. L. , Rashtchian A. . ( 1993; ). Dealing with contamination: enzymatic control of carryover contamination in PCR. . PCR Methods Appl 3: (Suppl.), S10–S14.[PubMed] [CrossRef]
    [Google Scholar]
  9. Health Protection Agency ( 2009a; ). One step influenza multiplex real-time RT-PCR National Standard Method SOP VSOP 50. .
  10. Health Protection Agency ( 2009b; ). European Real-Time RT-PCR for Influenza A H5 viruses. . National Standard Method VSOP 46 Issue 2 http://www.hpa-standardmethods.org.uk/pdf_sops.asp
  11. Health Protection Agency ( 2009c; ). Real-time RT-PCR (Taqman) for influenza A H5 viruses. . National Standard Method VSOP 41 Issue 5 http://www.hpa-standardmethods.org.uk/pdf_sops.asp
  12. Health Protection Agency ( 2009d; ). Swine-lineage influenza A H1 specific fast real-time PCR. . National Standard Method VSOP 29 Issue 5 http://www.hpa-standardmethods.org.uk/pdf_sops.asp
  13. Health Protection Agency ( 2009e; ). Swine-lineage influenza A N1 fast real-time confirmatory PCR assay. . National Standard Method VSOP 48 Issue1 http://www.hpa-standardmethods.org.uk/pdf_sops.asp
  14. Julkunen I. , Melén K. , Nyqvist M. , Pirhonen J. , Sareneva T. , Matikainen S. . ( 2000; ). Inflammatory responses in influenza A virus infection. . Vaccine 19: (Suppl. 1), S32–S37. [CrossRef] [PubMed]
    [Google Scholar]
  15. Leber A. . & other authors ( 2010; ). Vaccines shown to contain PCR-detectable DNA include Pentacel®, Daptacel®, and Adacel®. Detection of Bordetella pertussis DNA in acellular vaccines and in environmental samples from pediatric physician offices. . Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC): Boston, USA.
    [Google Scholar]
  16. McCaughey C. . ( 2010; ). Influenza: a virus of our times. . Ulster Med J 79:, 46–51.[PubMed]
    [Google Scholar]
  17. Meader E. , Waters J. , Sillis M. . ( 2008; ). Chlamydia trachomatis RNA in the environment: is there potential for false-positive nucleic acid amplification test results?. Sex Transm Infect 84:, 107–110. [CrossRef] [PubMed]
    [Google Scholar]
  18. Speers D. J. . ( 2006; ). Clinical applications of molecular biology for infectious diseases. . Clin Biochem Rev 27:, 39–51.[PubMed]
    [Google Scholar]
  19. van Elden L. J. R. , Nijhuis M. , Schipper P. , Schuurman R. , van Loon A. M. . ( 2001; ). Simultaneous detection of influenza viruses A and B using real-time quantitative PCR. . J Clin Microbiol 39:, 196–200. [CrossRef] [PubMed]
    [Google Scholar]
  20. WHO ( 2007; ). Recommendations and laboratory procedures for detection of avian influenza A (H5N1) viruses in specimens from suspected human cases. . Available from http://www.who.int/influenza/resources/documents/h5n1_laboratory_procedures/en/index.html
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.036178-0
Loading
/content/journal/jmm/10.1099/jmm.0.036178-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error