Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis

N K Fry; Jette Marie 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

Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis

N K Fry, Jette Marie 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

Department of Immunology and Microbiology

49 Citations (Scopus)

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.

Original language	English
Journal	European Journal of Clinical Microbiology & Infectious Diseases
Volume	19
Issue number	10
Pages (from-to)	773-80
Number of pages	8
ISSN	0934-9723
Publication status	Published - 2000

Keywords

DNA, Bacterial
Europe
Genotype
Humans
Legionella pneumophila
Legionnaires' Disease
Multicenter Studies as Topic
Polymorphism, Genetic
Polymorphism, Restriction Fragment Length
Reproducibility of Results
Serotyping

Cite this

Fry, N. K., Bangsborg, J. M., Bernander, S., Etienne, J., Forsblom, B., Gaia, V., Hasenberger, P., Lindsay, D., Papoutsi, A., Pelaz, C., Struelens, M., Uldum, S. A., Visca, P., & Harrison, T. G. (2000). Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis. European Journal of Clinical Microbiology & Infectious Diseases, 19(10), 773-80.

Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis. / Fry, N K; Bangsborg, Jette Marie; Bernander, S et al.

In: European Journal of Clinical Microbiology & Infectious Diseases, Vol. 19, No. 10, 2000, p. 773-80.

Research output: Contribution to journal › Journal article › Research › peer-review

Fry, NK, Bangsborg, JM, Bernander, S, Etienne, J, Forsblom, B, Gaia, V, Hasenberger, P, Lindsay, D, Papoutsi, A, Pelaz, C, Struelens, M, Uldum, SA, Visca, P & Harrison, TG 2000, 'Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis', European Journal of Clinical Microbiology & Infectious Diseases, vol. 19, no. 10, pp. 773-80.

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T1 - Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis

AU - Fry, N K

AU - Bangsborg, Jette Marie

AU - Bernander, S

AU - Etienne, J

AU - Forsblom, B

AU - Gaia, V

AU - Hasenberger, P

AU - Lindsay, D

AU - Papoutsi, A

AU - Pelaz, C

AU - Struelens, M

AU - Uldum, S A

AU - Visca, P

AU - Harrison, T G

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N2 - 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.

AB - 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.

KW - DNA, Bacterial

KW - Europe

KW - Genotype

KW - Humans

KW - Legionella pneumophila

KW - Legionnaires' Disease

KW - Multicenter Studies as Topic

KW - Polymorphism, Genetic

KW - Polymorphism, Restriction Fragment Length

KW - Reproducibility of Results

KW - Serotyping

M3 - Journal article

C2 - 11117642

SN - 0934-9723

VL - 19

SP - 773

EP - 780

JO - European Journal of Clinical Microbiology & Infectious Diseases

JF - European Journal of Clinical Microbiology & Infectious Diseases

IS - 10

ER -

Assessment of intercentre reproducibility and epidemiological concordance of Legionella pneumophila serogroup 1 genotyping by amplified fragment length polymorphism analysis

Abstract

Keywords

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