European Journal of Clinical Microbiology and Infectious Diseases

, Volume 19, Issue 10, pp 773–780

Assessment of Intercentre Reproducibility and Epidemiological Concordance of Legionella pneumophila Serogroup 1 Genotyping by Amplified Fragment Length Polymorphism Analysis

  • N. K. Fry
  • J. M. Bangsborg
  • S. Bernander
  • J. Etienne
  • B. Forsblom
  • V. Gaia
  • P. Hasenberger
  • D. Lindsay
  • A. Papoutsi
  • C. Pelaz
  • M. Struelens
  • S. A. Uldum
  • P. Visca
  • T. G. Harrison
Article

DOI: 10.1007/s100960000359

Cite this article as:
Fry, N., Bangsborg, J., Bernander, S. et al. EJCMID (2000) 19: 773. doi:10.1007/s100960000359

Abstract

 The aims of this work were to assess (i) the intercentre reproducibility and epidemiological concordance of amplified fragment length polymorphism analysis for epidemiological typing of Legionella pneumophila serogroup 1, and (ii) the suitability of the method for standardisation and implementation by members of the European Working Group on Legionella Infections. Fifty coded isolates comprising two panels of well-characterised strains, a “reproducibility” panel (n=20) and an “epidemiologically related” panel (n=30), were sent to 13 centres in 12 European countries. Analysis was undertaken in each centre following a previously determined standard protocol. Results were analysed by the participants, using gel analysis software where available, and submitted to the coordinating centre. The coordinating centre reanalysed all results visually and selected data-sets with gel analysis software. Data analysis by participants yielded reproducibility (R) values of 0.20–1.00 and epidemiological concordance (E) values of 0.11–1.00, with 6 to 34 types. Following visual analysis by the coordinating centre, R=0.78–1.00, and E=0.67–1.00, with 10–20 types. Analysis of three data-sets by the coordinating centre using gel analysis software yielded R=1.00 and E=1.00, with 12, 13 or 14 types. This method can be used as a simple, rapid screening tool for epidemiological typing of isolates of Legionella pneumophila serogroup 1. Results demonstrate that the method can be highly reproducible (R=1.00) and epidemiologically concordant (E=1.00), with good discrimination. The electropherograms generated are amenable to computer-aided analysis, but strict adherence to a previously defined laboratory protocol is required. Following designation of representative type strains and patterns, this method will be adopted by the European Working Group on Legionella Infections as the first internationally standardised typing method for use in the investigation of travel-associated Legionella infections.

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • N. K. Fry
    • 1
  • J. M. Bangsborg
    • 2
  • S. Bernander
    • 3
  • J. Etienne
    • 4
  • B. Forsblom
    • 5
  • V. Gaia
    • 6
  • P. Hasenberger
    • 7
  • D. Lindsay
    • 8
  • A. Papoutsi
    • 9
  • C. Pelaz
    • 10
  • M. Struelens
    • 11
  • S. A. Uldum
    • 12
  • P. Visca
    • 13
  • T. G. Harrison
    • 1
  1. 1.Respiratory and Systemic Infection Laboratory, Public Health Laboratory Service Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK e-mail: nfry@phls.nhs.ukGB
  2. 2.Department of Clinical Microbiology, Rigshospitalet, Copenhagen, DenmarkDK
  3. 3.Department of Clinical Microbiology, Karolinska Sjukhuset, Stockholm, SwedenSE
  4. 4.Centre National de Référence des Legionella et des Staphylocoques, Lyon, FranceFR
  5. 5.National Public Health Institute, Legionella Reference Laboratory, Helsinki, FinlandFI
  6. 6.Istituto Cantonale Batteriosierologico, Lugano, SwitzerlandCH
  7. 7.Federal Public Health Laboratory, Vienna, AustriaAT
  8. 8.Scottish Legionella Reference Laboratory, Stobhill Hospital, Glasgow, ScotlandGB
  9. 9.Department of Microbiology, School of Medicine, Aristotelian University of Thessaloniki, Thessaloniki, GreeceGR
  10. 10.Centro Nacional de Microbiologia, Majadahonda, Madrid, SpainES
  11. 11.Service de Microbiologie, Laboratoire de Référence des Legionella, Hopital Erasme, Brussels, BelgiumBE
  12. 12.Department of Respiratory Infections, Meningitis and Sexually Transmitted Diseases, Statens Serum Institut, Copenhagen, DenmarkDK
  13. 13.Unit of Special Pathogens, Istituto Superiore di Sanità, Rome, Italy Present address: Department of Biology, University of Rome III and Molecular Microbiology Unit, IRCCS “Lazzaro Spallanzani”, Rome, ItalyIT